Waste Biomass Valor
DOI 10.1007/s12649-017-9862-8
ORIGINAL PAPER
Current Treatment Technologies of Cheese Whey and Wastewater
by Greek Cheese Manufacturing Units and Potential Valorisation
Opportunities
K. Valta1 · P. Damala1 · E. Angeli1 · G. Antonopoulou2 · D. Malamis1 ·
K. J. Haralambous1
Received: 15 June 2016 / Accepted: 13 February 2017
© Springer Science+Business Media Dordrecht 2017
Abstract In the present study, recording and evaluation
of the current practices applied for the treatment of waste-
water and cheese whey, derived from ten cheese manufac-
turing units in Greece, was performed. Wastewater from
cheese manufacturing includes milk losses, wastewater
generated by the staff and by the washing of equipment,
floors and milk transferring vehicles. Based on the study,
cheese whey is separately collected from the cheese man-
ufacturing wastewater in almost all the Greek units under
study. Wastewater generated by Greek medium size units
is mainly treated through the extended aeration activated
sludge process, while wastewater derived from small size
units is either disposed in the sewage system or it is stored
until its transportation for further treatment. Moreover,
aiming at revealing the potential exploitation opportunities,
a comprehensive literature review on cheese whey valorisa-
tion technologies was also conducted. The literature review
revealed that cheese whey and whey components can be
used in a wide range of applications, mainly in the food
and nutraceutical industry, as they contain several benefi-
cial compounds that can be exploited. In this context, the
increasing consumer concerns related to health and well-
being have enhanced the acceptability and demand for
whey products, such as the whey protein concentrates and
isolates, the whey powder, the reduced lactose whey, etc.
Moreover, cheese whey can be used for the production of
* K. Valta
katvalta@gmail.com
1 trated milk whether or not sweetened; milk or cream in
School of Chemical Engineering, National Technical
University of Athens, 9 Iroon Polytechniou Str., Zographou
Campus, 15780 Athens, Greece
2
Institute of Chemical Engineering Sciences (ICE-HT/
FORTH), Stadiou, Platani, 26504 Patras, Greece
sustainable energy alternatives to fossil fuels, such as etha-
nol, hydrogen and biogas. Among them, biogas produc-
tion through anaerobic digestion could be a viable solution
since relevant economic analysis revealed that this technol-
ogy is a profitable and sustainable investment for a Greek
cheese whey factory.
Keywords Cheese manufacturing wastewater · Cheese
whey · Valorisation · Treatment · Greece · Economic
evaluation
Introduction
Globally, the dairy industry is one of the most important
processing sectors of the food and beverage industry. In
2010, Europe was the largest exporter of dairy products in
the world, accounting for approximately 13% of the food
and drink sector turnover in the EU [1]. The dairy sector
constitutes one of the most important players of the Greek
food and beverage industry. In 2012, 6% of the Greek food
and beverages companies were engaged in the dairy indus-
try [2]. Moreover, the dairy industry sector employed 12%
of the total number of employees contributing with 17 and
15% to the total turnover and value added of the food and
beverage sector respectively [2].
Dairy industry is related to the processing of the raw
milk into products such as fresh liquid milk, pasteurised,
sterilised, homogenised and/or ultraheat treated milk;
milk-based drinks; cream from milk; dried or concen-
solid form; butter; yoghurt; cheese and curd; whey; and
casein or lactose. In 2014, across the EU-28, the farms
produced approximately 1.648 billion tonnes of milk,
68.8 million tonnes of which were utilised to produce
13
Vol.:(0123456789)

5.5 million tonnes of cheese [3]. More than 300 different
kinds of cheese and other dairy products count millions
of fans all over the world and are officially registered and
protected as geographical indications or traditional spe-
cialties in EU-28 [4].
The basic principles of cheese making process are
common. However, many variations exist in all stages of
the process, resulting in the production of many differ-
ent varieties, often from the same factory [1]. Cheese is
produced by coagulation of the milk protein, casein, in a
way that traps milk solids and fat into a curd matrix [1].
Milk is mixed with a starter culture of bacteria and the
enzyme rennet, which catalyses casein coagulation [1].
In the sequel, the curd is cut into cubes and the mixture
is stirred slowly so as to collect as much protein as pos-
sible into the curd [1]. The cheese yield is approximately
10%, with the remaining 90% being a liquid by-product
called “whey” [1]. The liquid whey is separated and
drained from the curd. The curd is salted, pressed, cured,
and packaged as cheese [1].
In particular, according to Decision 97/80/EC: ‘whey
is a by-product obtained during the manufacture of
cheese or casein’. In the liquid phase, whey contains nat-
ural constituents (on average 4.8% lactose, 0.8% protein
and 0.2% fats by weight of the product) which remain
when the casein and the majority of the fat have been
removed from the milk [5]. Fifty years ago, cheese whey
(CW) was considered as wastewater from the processing
of cheese and it was mainly disposed along with waste-
water generated by the washing of equipment and floors
of the cheese manufacturing units. In many cases, this
stream was inappropriately disposed to the environment
without any prior treatment. As a result due to the high
organic load of CW serious environmental problems
were caused. However, it was later observed that whey
can be utilised for the production of white cheese e.g. for
“manouri”, “anthotyro”, “mizithra”, “giza”; and also uti-
lised as animal feed or for protein production. This new
direction in whey valorisation resulted in the opening of
a new market opportunity. Thus, nowadays food manu-
facturers use whey as a functional ingredient in food and
pharmaceutical applications, and as nutrient in dietetic
and health foods [3].
Although there are various studies and scientific pub-
lications related to the valorisation of CW worldwide, a
comprehensive study revealing opportunities by cheese
manufacturing wastewater and CW management for
Greece does not exist. The present study aims at explor-
ing potential valorisation opportunities from wastewater
and/or CW generated by Greek cheese manufacturing
units by taking into account the currently applied meth-
ods for the treatment of both waste streams.
13
Waste Biomass Valor
Methodology
In the present survey the term ‘wastewater’ is used to
describe the stream that is mainly derived from the clean-
ing of the equipment, the washing of the plant floor and of
vehicles for the transport of milk and from the staff. CW
is considered as a process by-product which is not treated
together with the wastewater a priori. The term ‘cheese
whey’ is mainly used to describe the primary whey gener-
ated during the production of cheese from raw milk while
the term ‘secondary whey’ refers to whey produced after
white cheese production. In order to shed light on oppor-
tunities by wastewater and cheese whey management in
Greece, the study was initially focused on recording waste-
water treatment applied to the Greek cheese manufacturing
sector. Following, international practices related to waste-
water and cheese whey treatment and valorisation were
reviewed aiming at revealing potential valorisation technol-
ogies that are unknown in the Greek context. Hence, both
current treatment practices in Greece and trends proposed
by the scientific community were studied and evaluated.
The same methodology has been also applied for explor-
ing waste and wastewater valorisation opportunities by the
Greek meat processing industry [6] and by the Greek olive
oil processing industry [7]. For the needs of the present
study, targeted questionnaires were distributed and face-to-
face interviews were conducted while further information
was taken from the Approvals of Environmental Impact
Assessment, officially issued by the Greek Ministry of
Environment and Energy. Collected data were related to
both the production process and the wastewater generation
and treatment. In total, ten small and medium cheese man-
ufacturing units operating in Greece were studied. They
were codified from 10.5–1 to 10.5–10; since numerical
code 10.5 characterizes the manufacture of dairy products
under the Statistical classification of economic activities in
the European Community (NACE Rev.2). Half of the units
are processing less than 15 tonnes of raw milk per day and
the rest are treating from 30 to 100 tonnes of raw milk per
day for the production of various types of hard, semi-hard,
white cheese, etc. Next, mapping, interpretation and assess-
ment of the collected information was implemented.
At the same time, a thorough literature review was con-
ducted. Both original research papers and review articles
related to the treatment and valorisation of cheese whey
were analyzed. In total, more than 300 abstracts’ publica-
tions were initially screened, out of which 55 were selected
for further study. From the selected papers, 42 were related
to the production of energy (biogas, hydrogen, ethanol,
butanol, electricity) and 13 to the valorisation of cheese
whey for the manufacture of valuable products. Addition-
ally, methods for the treatment of cheese whey are also
included, which mainly concern the anaerobic digestion
Waste Biomass Valor
process that results in the production of biogas. Results
obtained from the application of the methodology are pre-
sented in detail below.
Results and Discussion
Present Situation Related to Cheese Whey
and Wastewater Treatment and Valorisation
in the Greek Cheese Producing Industries
Industrial unit 10.5-1 is a small cheese manufacturing
unit with a capacity of 0.8 tonnes of milk per day which
operates 260 days per year. The installed power is equal
to 15.7 kW motor power and 15 kW thermal power. It is
estimated that 0.2 tonnes of cheese are produced, in a daily
basis. During the production process the milk is stored
in milk cooling tanks and is then transferred for filtering
purposes to a centrifugal separator. Filtered milk is mildly
heat-treated while stirring and is then cooled. After cool-
ing, the milk is transferred to tanks where rennet is added
and curd is produced. The curd is then separated from
whey and ripening of curd takes place and finally cheese is
maintained in a cooling chamber. Wastewater stream gen-
erated by the entire process includes whey and wastewater
produced after washing of the equipment and floors. Whey
is collected in separate tanks and is provided to a nearby
pig farm as feed while wastewater from the equipment and
floor washing, is directed to the sewage system.
Industrial unit 10.5-2 is a small cheese manufacturing
unit with a daily capacity of 1 tonne of milk per day, for the
production of “feta”, “mizithra”, yogurt and “kefalotiri”.
The installed power of the plant is 17.05 kW. In brief, the
production process includes the delivery and storage of the
milk; its pasteurisation and standardising processes; pro-
duction of feta cheese; and production of other dairy prod-
ucts such as “mizithra”, yogurt and “kefalotiri”. Wastewa-
ter which is generated from the whole production process,
equipment and floors washing are stored to tanks for a short
period and are then led to the wastewater treatment plant
(WWTP) of the municipality. Part from CW is used for the
production of “mizithra”. Finally, primary and secondary
cheese whey (700 kg per day) are delivered to a big farm as
feed. Industrial unit 10.5-3 is also a small cheese manufac-
turing unit with a daily capacity of 2 tonnes of milk for the
production of 0.5 tonnes white cheese per day. The power
is equal to 31 kW motor power and 6 kW thermal power.
Wastewater is generated by the washing of the equipment
and floors; and from the staff. Whey is collected in separate
tanks and is provided to a pig farm as feed, while wastewa-
ter is directed to a separate tank from where it is forwarded
for further treatment. Industrial unit 10.5-4 has a daily
capacity of processing 4.6 tonnes of milk from cows, sheep
and goats per day. The cheese-making unit produces some
traditional Greek cheeses i.e. “metsovone”, “metsovella”,
“graviera” and cheese made from goat milk. The installed
power is equal to 44 kW. For the needs of the process
approximately 20,670 m3 of water are consumed yearly.
Wastewater includes the wastewater generated by the staff;
water from the washing of the equipment, the floors and the
vehicles that transfer the milk; and milk losses from the
production process. Whey is separated and transferred
either to pig farms to be used as feed or as substrate of
anaerobic digesters, while the remaining wastewater is dis-
posed to the municipal sewage system. The anaerobic
digestion treatment is executed by an independent company
operating anaerobic digesters close to the industrial unit.
Wastewater other than whey is disposed to the municipal
sewage system. Industrial unit 10.5-5 is a cheese manufac-
turing unit with a daily capacity of 4.65 tonnes of milk per
day for the production of 1 tonne of cheese. The total
installed power is equal to 80 kW. The raw materials used
in the plant include milk from cows, sheep and goats; salt;
and rennet. In total 14 m3 of wastewater is produced in a
daily basis. Moreover, 3.57 tonnes of CW is generated in
each day, from which 3% is utilised for “mizithra” produc-
tion while the remainder is provided to a farm as a feed for
pigs. For the treatment of wastewater, activated sludge pro-
cess is applied through sequencing batch reactors (SBR).
The treated effluent is chlorinated and is disposed under-
ground. Industrial unit 10.5-6 is processing up to 15 tonnes
of raw milk per day with an installed power of the plant
being 135.48 kW. Cheese whey is co-treated with wastewa-
ter from the cleaning of the equipment and floors through
an attached growth aerobic process with trickling filters.
Treated effluent is disposed to the sewerage system. Indus-
trial unit 10.5-7 has a daily capacity of treating 30 tonnes
of milk from sheep and goats. In total, 10,000 m3 of water
are consumed to cover the needs of the unit. Thus, high
volumes of wastewater are generated including the washing
water (of the equipment, the floors and the vehicles that
transfer the milk); wastewater generated by the staff and
milk losses from the production process. Wastewater is
treated in a WWTP which operates at the premises of the
industrial unit, where preliminary, primary, secondary and
tertiary treatment is conducted. More particularly, the treat-
ment process includes screening, flow equalisation, coagu-
lation, air flotation, biological treatment with extended aer-
ation activated sludge process and finally, chlorination. A
portion of the biological sludge is recycled to the aeration
tank to maintain a high mixed-liquor suspended solids
level, while the remainder is removed from the process and
is sent to sludge processing where stabilisation and then
dewatering of the sludge in drying beds is carried out. CW
is collected and provided as animal feed, in a daily basis
while in the case that whey transportation cannot be
13

achieved in the same day, CW is kept to refrigerators.
Industrial unit 10.5-8 is a large cheese manufacturing unit
with a daily capacity of 40 tonnes of milk per day and a
total installed power of 292.75 kW (motor) and of 39 kW
(thermal). The water consumption is approximately
15,000 m3 per year. The raw materials used in the plant are
mainly the milk, salt and rennet. The main end-products are
cream and the traditional Greek cheeses “kasseri” and
“mizithra”. The milk is received by local producers and
after quality control is transferred into tanks. In the sequel,
milk is cooled at temperature below 6 °C with the aid of a
cooling device (chiller), to be used in the production pro-
cess of “kasseri” or whey or “mizithra”. The milk from
these tanks is pre-heated in a temperature range of 32 °C to
37 °C and then is directed to a cream separator machine.
The centrifugation process is applied in order to remove
any impurities such as plant fibers, soil, etc, from the milk.
The fat, which is also separated, is then mixed again with
the skimmed milk. For the production of “kasseri” cheese,
the milk is introduced in tanks and rennet is also added to
cause milk coagulation. The mixture stays in the tanks for
10–20 min in temperatures between 32 to 37 °C. Following,
the curd is cut into small pieces and then is heated
(32–37 °C) while is agitated and the mixture stays for
10–15 min. The heating step helps to separate the whey
from the curd. Following, the curd is extracted and is then
pressed in order to obtain the desired shape. Afterwards the
products are left for ripening for 24–48 h at 14–18 °C in a
slightly acidic aqueous environment (pH 5.1–5.4). After-
wards cutting to the desired dimensions is implemented
and the curd enters the machine for making the “kasseri”.
There, the cheese mass is heated to 72 °C up 78 °C and salt
(about 2% by weight) is added. At the final stage inside the
“kasseri” machine, moulding is implemented at ambient
temperature. Then the first ripening, which lasts for 7 days,
is executed. The products after ripening are rinsed with
water, dried and covered with paraffin. In the last step, the
“kasseri” is cut into pieces and is stored in coolers (2–4 °C)
for the final ripening for 3 months. As in the case of “kas-
seri”, for the production of “mizithra” cheese, the milk,
which is kept in cooling conditions, is entering the cream
separator in order to remove the cream from skimmed milk.
The cream is then packed and is given for sale. Then the
milk enters the tanks to be heat- treated at 85–90 °C, for
5 min. The formation of the curd takes place and after
10 min the curd is removed and is placed to cheesecloths
where salt is added. CW is also removed. Drain takes place
twice and cheesecloth is removed. Then crushing and pack-
ing into containers is implemented and the final product is
refrigerated to 4 °C, until the placement of the product to
the market for sale. From the operation of the unit, waste-
water (100 m3/d) and whey (30 m3/d) are produced. CW is
separately collected and is provided as feed in big farms.
13
Waste Biomass Valor
Wastewater is mainly generated by washing and cleaning of
the production and the packaging processes, as well as by
the cleaning of the vehicles that transfer the raw material
(milk).Wastewater is treated in a WWTP at the premises of
the unit. During its treatment process, screening of the
wastewater initially takes place and then it is directed into a
flow equalization tank. In the sequel, coagulation and dis-
solved air flotation (DAF) take place and finally, biological
treatment by extended aeration of the activated sludge is
carried out. Secondary treated effluent is subjected to a
chlorination tank and is then disposed to a stream which
ends to a river. Sludge, generated by the biological treat-
ment process, is treated in drying beds (180 m2). Dewa-
tered sludge has a total solids content of approximately
80%. Industrial unit 10.5-9 is a large cheese manufacturing
unit with a daily capacity of 62 tonnes of milk per day. The
motor power of the plant is 710.66 kW which also includes
power needs of the WWTP. Additional 193.63 kW power
for the auxiliary equipment is needed. The products that are
manufactured are: feta cheese, “mizithra”, “giza”, butter,
“anthotyros”, semi hard cheese and yogurt. Wastewater is
separately treated in a WWTP including DAF, anaerobic
digestion in an upflow anaerobic sludge blanket (UASB)
reactor and aerobic treatment through extended aeration of
the activated sludge. The excess of sludge is separately
treated in a filter press. Part of produced biogas is utilised
to cover the needs of the unit, while the remainder is burnt.
Whey (6486 tonnes/y) is collected and given to third par-
ties for protein production. Industrial unit 10.5–10 is a
large cheese manufacturing unit with a daily capacity of
100 tonnes of milk per day for the production of “feta”,
hard cheese and different types of “mizithra”. The installed
power of the plant is 733.11 kW. Water consumption is
approximately 75.600 m3 per year. Wastewater generated
by the washing of the equipment along with the respective
produced by the staff is co-treated to the WWTP of the
unit. The WWTP has been designed for treating310 m3 per
day and includes screening, flow equalisation, biological
treatment with extended aeration activated sludge process,
settling and chlorination. In addition, sludge dewatering is
carried out, through sludge filter press. From the other side,
there is no treatment process for CW, which is collected
and provided as animal feed. Currently applied wastewater
treatment methods and ways for disposal of the treated
effluent of the Greek dairy industrial units under study, are
summarised in Table 1.
As it is noted above, CW is separately collected in
almost all units studied. Only wastewater and CW gener-
ated in industrial unit 10.5–6 are co-treated as a single
wastewater stream. CW is used either for the production of
white cheeses by many of the industrial units studied or it
is directly provided as animal feed for pigs. The use of CW
as animal feed seems to be the most economically feasible
Waste Biomass Valor

Table 1  Currently applied wastewater treatment methods and ways of disposal of the treated effluent of the Greek dairy industrial units under
study
Code 10.5 Capacity, Products Wastewater (other than whey) treatment Whey treatment & valorisation
tonnes of milk method applied & disposal of the treated
per day effluent

1 0.8 Cheese Treatment: collection and disposal to septic Valorisation: collection and provision as
tanks animal feed for pigs
Disposal: to the sewerage system
2 1 Feta cheese Treatment: collection to tanks and then led Valorisation (0.7 t/d): production of mizithra
Mizithra to the municipal WWTP and the excess is provided as animal feed
Yogurt Disposal: na*
Kefalotiri
3 2 Cheese Treatment: collection to tanks and then led Valorisation: collection and provision as
for further treatment animal feed for pigs
Disposal: na*
4 4.6 Metsovone Treatment:- Valorisation: collection and provision as
Metsovella Disposal: to the municipal sewerage animal feed for pigs and as feedstock to an
Graviera system anaerobic digestion treatment plant outside
Cheese from goat milk the premises of the unit
5 4.65 Cheese Treatment (14 m3/d): SBR Valorisation (3.57 t/d): 3% used for the
Disposal: underground production of mizithra and the excess is
provided as animal feed for pigs
6 15 Cheese Treatment:- Cheese whey is co-treated with wastewater from the cleaning of the equip-
ment and floors through trickling filters
Disposal: to the municipal sewerage system
7 30 Cheese Treatment Valorisation: collection and provision as
Screening animal feed
Flow equalisation
Oil and grease removal
Biological treatment by extended aeration
activated sludge process
Chlorination of the treated effluent
Collection of the sludge to the stabilisa-
tion tank where sludge stabilisation
under continuous aeration takes place
Disposal: Surface water recipient (river)
8 40 Kasseri Treatment (100 m3/d) Valorisation (30 t/d): collection and provi-
Cream Biological treatment sion as animal feed for pigs
Mizithra The sludge is treated in drying beds
Disposal: Surface water recipient (river)
9 62 Feta cheese Treatment Valorisation (6486 t/year): collection and
Mizithra Flow equalisation provision for protein production
Giza pH control
Butter Anaerobic process through the use of
Anthotyro UASB reactors
Semi hard cheese Biological treatment with extended aera-
Yogurt tion activated sludge process
Settling
10 100 Feta cheese Treatment (310 m3/d) Valorisation: collection and provision as
Hard cheese Screening animal feed
Mizithra Flow equalisation
Biological treatment with extended aera-
tion activated sludge process
Settling
Chlorination

*na not available

option, since pig farms are located close to the cheese man- production of proteins was also recorded, as the currently
ufacturing units which in turns makes daily whey trans- applied method by industrial unit 10.5–10. This unit is a
port easy, even in small quantities. CW valorisation for the medium size unit resulting in the generation of large CW

13

volumes, which might explain the specific CW use. Finally,
industrial unit 10.5–4 provides the excess of CW either for
animal feeding purposes or as a feedstock in an anaero-
bic treatment unit. Different valorisation routes for cheese
whey in the Greek units studied are presented in Fig. 1.
Apart from CW, wastewater is also generated by the
staff; the washing of the equipment, the floors and the vehi-
cles that transfer the milk; including also milk due to milk
losses from the production process. From the Table 1, it
was observed that the existence of more technologically
advanced wastewater treatment systems was related to the
capacity of the cheese manufacturing unit. For instance
wastewater generated by the industrial units 10.5–1 (0.8
tpd) and 10.5–4 (4.6 tpd) is directed to sewerage system.
Moreover, wastewater resulting for the manufacturing of
cheese by industrial units 10.5–2 (1 tpd) and 10.5–3 (2
tpd) is stored to tanks for a short period and is then led to
central WWTP. Industrial units 10.5–5, 10.5–6, 10.5–7,
10.5–8, 10.5–9 and 10.5–10 operate their own WWTP at
their premises. The activated sludge process is applied for
the treatment of the generated wastewater at the majority of
the industrial units 10.5–5 use an SBR 10.5–7, 10.5–8 and
10.5–10 use extended aeration. At industrial unit 10.5–9,
Fig. 1  Cheese whey manage-
ment in the studied Greek
cheese manufacturing units
13
Waste Biomass Valor
the generated wastewater is treated in an UASB reactor,
followed by an extended aeration activated sludge process.
Finally in industrial unit 10.5–6, the wastewater and CW
are co-treated through the attached growth aerobic process
with the application of trickling filters. The applied man-
agement for wastewater produced by the operation of the
Greek units studied are presented in Fig. 2.
Potential Valorisation and Treatment Opportunities
for Wastewater and CW Coming from Greek Cheese
Producing Industries
Anaerobic Digestion Treatment
Based on the literature review concerning whey and waste-
water treatment and valorisation systems, the anaerobic
digestion (AD) process seems to be the most studied tech-
nology. Thus, Saddoud et al. [8] applied two-stage anaero-
bic digestion of CW consisting of a stirred tank acidogenic
reactor followed by a CSTR methanogenic reactor. The
second reactor was coupled to a membrane filtration sys-
tem, in order to remove the soluble solids from the effluent.
According to the results of that study, quality characteristics
Waste Biomass Valor
Fig. 2  Applied management for wastewater produced by the operation of the studied Greek cheese manufacturing units
of the permeate were satisfactory and below the acceptable
standards for the disposal through public drains in Tunisia
(chemical oxygen demand (COD) < 90 mg/L, biochemi-
cal oxygen demand (BOD) < 30 mg/L and total suspended
solids (TSS) < 90 mg/L). Through the two-stage anaero-
bic digestion process, high removal efficiency values were
observed i.e. 98.5, 99 and 100% for COD, ­BOD5 and TSS,
respectively. Moreover, the daily biogas production rate
exceeded 10 times the reactor volume and the methane con-
tent in the biogas was greater than 70%, showing the high
performance of the process. Diamantis et al. [9] studied the
anaerobic digestion of original CW which was diluted with
wastewater, either in an UASB reactor or in a two phase
AD system, consisting of an acidogenic CSTR followed by
a UASB reactor, with and without biomass recirculation.
Results clearly indicated that the two-stage configuration
was superior in terms of process efficiency and methane
recovery when compared to the single-stage system, while
the enhanced acidification process resulted in optimum
operational and performance features, achieving up to 90%
COD removal and high methane yields [9].Gannoun et al.
[10] investigated the effect of biological pretreatment on
the AD of CW through an upflow anaerobic filter (UAF)
operating at various organic loading rates (OLR) and
hydraulic retention times (HRT). The pretreatment step was
conducted with Lactobacillus paracasei which was growth
on CW, conducting lactose fermentation into lactic acid.
The organic matter was also precipitated with lime which
resulted in 50% COD reduction and 60% TSS reduction.
The pretreated feedstock was then introduced into the UAF
in which the average COD removal achieved was 80–90%.
The valorisation of different types of agroindustrial
wastes through anaerobic co-digestion has been addressed
by many authors as a promising method for achieving both
waste treatment and renewable energy production. In par-
ticular, Rico et al. [11] applied co-digestion process using
one-stage UASB reactor with an external settler and efflu-
ent recycling for alkalinity supplementation. The process
demonstrated stable operation when fed with a CW frac-
tion of 75% and operated with an applied OLR of 19.4 kg
COD ­m− 3 ­d− 1, achieving 94.7% efficiency removal and a
methane production rate of 6.4 m3 ­CH4 ­m− 3 ­d− 1. Critical
biomass washout was observed when the CW fraction in
the feed increased to 85%. In the same study was found that
the use of lower CW fractions e.g. 60%, under high OLR
(28.7 kg COD ­m− 3 ­d− 1) and low HRT values (1.3 d) a sta-
ble operation was also observed achieving a removal effi-
ciency of 95.1% and the highest methane production rate
of 9.5 m3 ­CH4 ­m− 3 ­d− 1. Based on the authors this method
constitutes an interesting alternative for the co-management
of CW and manure.
Bertin et al. [12] also studied the co-digestion of CW
with cattle manure but using a two-stage configuration pro-
cess. The results showed that the addition of cattle manure
improved the anaerobic digestion process performance,
while the application of a two stage process exhibited
higher methane yields, compared to the respective of the
one stage system. Comino et al. [13] studied the feasibility
13

of co-digestion of CW with cattle slurry in a dairy farm.
The results of that study showed that production of methane
from a mix of 50:50 (v/v) of both feedstocks, without pH
adjustment through chemicals, was possible exhibiting a
methane yield of 343.43 L ­ CH4/kgVS for OLR of 2.65 gVS/
L/d. In the same line, Kavacik et al. [14] explored the pos-
sibility of biogas production from the co-digestion of CW
with dairy manure either at 25 or 34 °C. The experimen-
tal results obtained verified that a co-treatment of different
waste types like manure and CW gives the possibility of
treating a waste which cannot be successfully treated sepa-
rately. Dareioti and Kornaros [15] had also investigated the
potential of mesophilic (37 °C) anaerobic co-digestion of a
pretreated mixture of ensiled sorghum, CW and liquid cow
manure (55:40:5, v/v/v) in a two-stage CSTR system. The
study was focused on the investigation of the HRT effect on
two different processes where at the first acidogenic step,
biohydrogen production is carried out, while at the sec-
ond stage methane production is conducted. According to
the authors, the highest H­ 2 production rate and yield were
observed at an HRT of 0.5 d during acidogenic stage, while
in methanogenic stage, the highest ­CH4 productivity was
observed at the HRT of 16 d. At an HRT of 12 d instability
of the methanogenic reactor occurred [15].
Anaerobic treatment may also be combined with aero-
bic processes. Thus, in the study of Frigon et al. [16] whey
wastewater was treated through an anaerobic digestion
step, followed by a sequential aerobic treatment step, under
low oxygen concentration. Both processes were conducted
sequentially in the same vessel, by changing the cycle
times in the same digester. An increase in either OLR or of
the total cycle time (days) resulted in an increase in COD
removal efficiency, reaching its maximum value of 98% at
0.78 g COD/L/d and a cycle time of 4 d. A second set of
experiments in the frames of the same study showed that
an improvement in the compartmentalization of the reac-
tor where the anaerobic and aerobic biomasses are con-
tained, is necessary, in order to enhance the degradation of
whey wastewater through sequential anaerobic and aerobic
processes.
Membrane Technologies
Another technology system that has been utilized for the
prevention of environmental pollution caused from whey
streams, could be the recovery of water and whey powder
from whey. This technique was investigated by Aydiner
et al. [17], who performed a techno-economical evalua-
tion of a CW treatment and valorization process, consist-
ing of alternative steps such as ultrafiltration (UF), reverse
osmosis (RO), forward osmosis (FO) and thermolysis (T).
In this context, three case studies were examined, includ-
ing UF/RO, FO/RO and FO/T/RO processes steps. Among
13
Waste Biomass Valor
the different alternative schemes tested, FO/RO with NaCl
draw solution, exhibited the most satisfactory results, pre-
senting the highest water recovery (77.4%) and whey pow-
der production (98.9 entity/year), along with a net present
value (NPV) of $12,347,000 and an inevitable result of
0.8 years payback lower than that of UF/RO.
Other Cheese Whey Wastewater Treatment Process
Finally, Prazeres et al. [18] utilized a Fenton-like oxida-
tion system for the treatment of CW wastewater, which
had been previously processed either through a coagula-
tion-flocculation step with F
­ eCl3, or through a sedimenta-
tion step with Ca(OH)2 and then an aerobic digestion step.
In the first case, a COD removal efficiency of 80% was
achieved and—depending on the initial concentration of
the reagent—the low retention times may result in almost
complete COD conversion efficiency, within few minutes.
Moreover, Ca(OH)2 addition enhanced the removal of
Fe(III) from the oxidation treatment, while an increase in
pH from 4.5 to 7.5 led to satisfactory settling properties.
Production of Valuable Products and Compounds
From a valorisation point of view, apart from energy pro-
duction, cheese whey can also result in the production of
valuable products and compounds. In particular, based
on the literature, CW can be considered as a valuable by-
product that can be valorized in the food and pharmaceuti-
cal industry, as it contains more than half of the solids pre-
sent in the original whole milk, including whey proteins,
lactose, water-soluble vitamins and minerals [18]. Accord-
ing to Mollea et al. [19], cheese whey management can be
accomplished via two main options: either by the recovery
of valuable compounds (e.g. proteins and lactose) with the
application of suitable technologies, or by the application of
fermentation processes in order to obtain value added prod-
ucts, such as organic acids, single cell proteins and oils,
biopolymers (enzymes, polyhydroxyalkanoates, etc.) and
bacteriocins [20]. In addition to this, when whey perme-
ate (obtained from the ultrafiltration step) is used as a fer-
mentation medium, both management options are applied.
Thus, depending on the processing technique that is used,
many types of whey products can be produced, namely, the
sweet and acid whey powder, the reduced lactose whey,
the demineralized whey, the whey protein concentrate
(WPC) (with varying protein contents), the whey protein
isolate (WPI), the mineral-concentrated whey (reduced
lactose whey), as well as lactoferrin, lactoperoxidase, gly-
comacropeptide and dairy minerals, such as calcium [21].
Palatnik et al. [22] used microfiltration and tangential ultra-
filtration in order to purify and concentrate caprine cheese
whey, reaching a protein concentration of 91.4/100 g.
Waste Biomass Valor
The concentrate, which was freeze-dried and character-
ized, showed good emulsifying properties, and high water
and oil holding capacity. The functional properties of the
caprine whey proteins were pH-dependent and their rheo-
logical behavior indicated that the whey concentrate can
be used in formulations that require gelation. In general,
the product developed presented similar physicochemical
characteristics to commercial dressings with regard to the
pH, ashes, viscosity, texture and color. The functionality of
caprine WPC was also investigated by Sanmartin et al. [23]
through the following consecutive processing methods:
thermocalcic precipitation, ultrafiltration/diafiltration and
lyophilisation of the retentate. Among the functional prop-
erties examined, only the solubility, emulsifying stability
and emulsifying capacity of the concentrates were higher
than those in commercial bovine WPC. Galanakis et al.
[24] investigated the separation and recovery of compounds
rich in proteins and sugars from whey substrates, which
were obtained during the processing of “Halloumi” cheese.
Experiments of ultrafiltration were conducted under con-
stant temperature, circulation flux and several transmem-
brane pressures. Two approaches were examined, the first
using a 100 kDa-polysulphone barrier and the second by
combining 20 kDa-polysulphone and 2 kDa-polyethersul-
phone membranes. Based on their findings, the combined
treatment optimizes protein recovery (87–90%) in the con-
centrate stream—despite the weaker separation with sug-
ars (39–32%)—and the recovered materials could be used
as valuable ingredients in yogurt, dairy desserts and meat
products. Folmer Corrêa et al. [25] examined the process
of enzymatic proteolysis for the release of bioactive pep-
tides. More specifically, a protease preparation from Bacil-
lus sp. was utilised for the hydrolysis of sheep CW and the
hydrolysates were evaluated for their antioxidant and angi-
otensin I-converting enzyme (ACE)-inhibitory activities.
During hydrolysis, soluble protein and free amino acids
increased for up to 4 h. Similarly, the antioxidant activity
of hydrolysates increased 3.2-fold, from 0 (15.9%) to 6 h
of hydrolysis (51.3%). As previously mentioned, whey can
also serve as a food-grade fermentation medium, for the
cultivation of producer strains and the production of bacte-
riocins, which can be used for food bio-preservation. In this
context, Kumari Garsa et al. [26] examined several pedi-
ocin producing strains, in terms of their capability to pro-
duce b-galactosidase, which can be used for the hydrolysis
of lactose. The authors conducted their experiments using
response surface methodology experimental design, which
was found to be a powerful tool in optimizing pediocin
PA-1 production. Their results indicated that pediocin PA-1
production is strongly affected by fermentation conditions
and CW can be used -at industrial scale- as a fermentation
medium for the production of food-grade biopreservatives.
Moreover, Watanabe et al. [27] investigated the effects of
soybean oil on the production of an enzyme (PaE) with
plastic-degrading activity by P. antarctica growing on
lactose or CW. Hence, when soybean oil was used as the
sole carbon sourcethe PaE activity was very low (0.03 U/
mL) but increased at 1.79 U/mL when a limited amount of
soybean oil (under 0.5%) was combined with a relatively
high concentration of lactose (6%). Besides, fermentation
with lactose fed-batch cultivation combined with periodic
soybean oil addition resulted in 14.6 U/mL of PaE, after
a 5-days cultivation, while when the lactose was replaced
with CW, PaE production was 10.8 U/mL after a 3-day cul-
tivation period. Galvão et al. [28] examined different con-
ditions of sequential hydrolyses of whey proteins in order
to produce protein hydrolysates with low Phenylalanine
(Phe) contents. For this purpose, trypsin, chymotrypsin and
carboxypeptidase A (CPA) were immobilized/stabilized on
special gel matrices and Phe was removed from the hydro-
lysates by the action of CPA, thus providing an adequate
source of proteins for phenylketonuria (PKU) patients. In
this context, proteins were sequentially hydrolyzed to dif-
ferent degrees of hydrolysis with trypsin and chymotrypsin
employing different enzyme/substrate ratios. The best
results were obtained using chymotrypsin directly to hydro-
lyze whey proteins and, based on the findings, a kinetic
model for the hydrolysis of CW proteins with immobilized
chymotrypsin was proposed, for high-load derivatives.
Sugai et al. [29] investigated yeast strains from several iso-
lates obtained from raw milk and milk products in terms
of their ability to produce glucosylceramide from CW.
The majority of strains that had accumulated a detectable
amount of glucosylceramide were identified as Kluyvero-
myces lactis var. lactis. Depending on the origin of K. lac-
tis var. lactis strains, variations in the quantity of gluco-
sylceramide and steryl glucoside produced, were detected
[29].
Following, two studies regarding the valorization of CW
for the production of other cheese products, will be ana-
lysed. Dimitrellou et al. [30] evaluated the performance of
free and immobilized Lactobacillus casei ATCC 393 on
whey protein as starter cultures for the production of pro-
biotic of the Feta-type cheese. This was achieved, firstly, by
ensuring that the probiotic strains can provide health bene-
fits at the time of consumption, secondly, by improving the
quality characteristics of the new products, and, finally, by
extending the products’ life by suppressing growth of spoil-
age and pathogenic bacteria. Thus, based on the authors,
whey protein can be used for the production of probiotic
foods as well as other highly added-value products. Finally,
Di Cagno et al. [31] produced low-fat Caciotta-type
cheeses testing the performance of several ingredients (par-
tially skim milk, microparticulated WPC, exopolysaccha-
rides (EPS)-producing Streptococcus thermophilus ST446,
as well as Lactobacillus plantarum (LP) and Lactobacillus
13

rhamnosus (LRA) as adjunct cultures) which were used
alone or in combination with each other. The experiments
revealed that microparticulated WPC increased the yield
of low-fat cheese under increased moisture content, while
EPS exhibited satisfactory results, especially in regard to
the enhancement of texture characteristics. Moreover, the
adjunct cultures increased the content in volatile compo-
nents (e.g. diacetyl and acetic acid) thus strengthening the
market acceptance of the low-fat Caciotta-type cheese.
Energy Production
Due to its high content in biodegradable carbohydrates,
CW may also constitute a promising feedstock for fer-
mentative hydrogen [32–48], ethanol [49–58] and butanol
[59] production, as well as for the production of electricity
through microbial cells [60–62]. All of these options can be
considered as sustainable energy alternatives to fossil fuels,
hence, a great number of studies found in the literature is
focused on this direction.
Hydrogen Production
More specifically, Reyes et al. [32] studied the effect of pH,
OLR and HRT οn inhibition of the methanogenic activity
in a hydrogenogenic UASB reactor treating CW. This work
demonstrated that it was possible to control the methane
activity in a hydrogen producing reactor using operational
strategies such as reduction of pH and HRT, organic shock
loads and repeated biomass heat treatment. Gomez-Romero
et al. [33] studied the production of biohydrogen by co-
digestion of CW with vegetables fruit waste. Results dem-
onstrated that the co-digestion of both substrates improves
biohydrogen production process, due to a better nutrient
balance and improvement of the system’s buffering capac-
ity. Vazquez et al. [34] investigated the production of con-
tinuous biohydrogen using powder solution CW, under dif-
ferent HRTs (10, 6 and 4 h) and OLRs (92.4, 115.5, 138.6
and 184.4 g lactose /L/d− 1) values. The experimental
results showed that the highest volumetric hydrogen pro-
duction rate (VHPR) was achieved at the HRT of 6 h, at all
OLRs which were tested. The highest VHPR (46.61 mmol
­H2/ L/ ­h− 1) and hydrogen molar yield of 2.8 mol ­H2/mol
lactose were found at an OLR of 138.6 g lactose/L/d, while
the lowest VHPR occurred at an OLR of 184.4 g lactose/ L/
d. Castello et al. [35] examined the applicability of dark
fermentation for hydrogen production in a UASB reactor
fed with raw and unsterilized CW, and the effect of OLR
increase, in hydrogen production performance. The results
showed that hydrogen can be produced but in low amounts.
For the highest loading rate tested (20 g COD / L/ d),
hydrogen production was 122 mL ­H2 / L/ d. It was reported
that operational pH value, is crucial parameter to promote
13
Waste Biomass Valor
­H2 production, but it is not sufficient to control and sup-
press methane production. Moreover it was found that the
biomass appeared to acclimatize to the low pH value, as
methanogenesis was restored even at pH 5. Venetsaneas
et al. [36] investigated the possibility of biological hydro-
gen production from CW and subsequent methane genera-
tion from the effluent of the hydrogenogenic process. Mes-
ophilic fermentative hydrogen production from undiluted
CW was investigated at an HRT of 24 h. pH culture was
maintained at a constant value of 5.2 either through alkalin-
ity addition or through an automatic pH controller. With
alkalinity addition, hydrogen production rate was
2.9 ± 0.2 L/ Lreactor/ d, while the yield of hydrogen pro-
duced was approximately 0.78 ± 0.05 mol H ­ 2/mol glucose
consumed. This yield corresponded to the production of
2.9 L of ­ H2/L CW. At pH controlling experiment, the
hydrogen production rate was 1.9 ± 0.1 L Lreactor/d, while
the yield of hydrogen was 0.61 ± 0.04 mol ­H2/mol glucose
consumed, corresponding to the production of 1.9 L of
­H2/L CW. Further conversion of the effluent of the hydrog-
enogenic reactor to methane was conducted in a continuous
anaerobic digester, operating at an HRT of 20 d in meso-
philic conditions. Approximately 1 L ­CH4/d was produced,
corresponding to a yield of 6.7 L ­CH4/L of influent. Taking
into account that a high COD removal was reported (>95%)
it was showed that CW could be efficiently used for hydro-
gen and methane production, in a two-stage process. In the
same line, Antonopoulou et al. [37] studied the combined
hydrogen and methane production from undiluted CW in a
two-stage process using a CSTR acidogenic reactor at the
first stage and a high-rate periodic anaerobic baffled reactor
(PABR) at the second methanogenic stage. It was shown
that continuous fermentative hydrogen production from
CW at an HRT of 24 h led to a hydrogen yield of 0.9 mol /
mol of glucose, corresponding to 2.49 L ­ H2/L of CW.
Regarding the second stage, the higher methane production
rate was obtained at an HRT of 4.4 days being 7.06 ± 0.29 L
L/day [37]. Fernández et al. [38] studied the application of
a packed bed reactor (polyurethane foam was applied as
support material) for the production of hydrogen from CW
permeate, focusing on the effect of OLR. Based on the
results, the decrease in OLR from 18.8 to 6.3 g COD/ L/ d
resulted in a significant drop in hydrogen production. The
obtained results revealed the presence of non-H2 producers,
Sporolactobacillus sp. and Prevotella, that might have
competed for substrate consumption and thus negatively
affected the prevalence of ­H2 producers microorganisms.
Despite that, hydrogen generation has been detected during
the entire experiment. In particular, hydrogen production
was favoured when feeding was interrupted. Moreover, H ­2
production was increased when higher OLR was applied,
while also maintenance of the pH of the feeding solution at
a value of 7.5 was beneficial for the process. Based on the
Waste Biomass Valor
specific research, operational conditions and possible pre-
treatment methods should be further studied so as to
increase hydrogen production. Seo et al. [39] developed a
pretreatment method for enhancing hydrogen production
from CW through a simple hydrodynamic cavitation (HC)
system under alkaline conditions. The proposed system
provided both the sonication effect but also an increase in
the temperature of the solution. Applying the hydrody-
namic cavitation pretreatment for 15 min, a higher hydro-
gen yield (1.89 mol ­H2/mol lactose) was achieved comper-
ing to that achieved with other conventional pretreatment
methods (e.g. heating and sonication). The efficiency of the
method was improved when combined with alkaline condi-
tions. In this way, the amount of solubilised nutrient was
greatly increased and methanogens were completely
removed, leading to H ­ 2 gas of higher purity (48%) and a
yield of 3.30 mol ­H2/mol lactose. The authors claimed that
the HC pretreatment under alkaline conditions can be used
to efficiently pretreat organic-rich wastewaters, including
CW, without any scale-up problem since is an excellent
pretreatment option for ­H2 production. Moreno et al. [40]
explored the potential of a two-stage process (fermentative
and biocatalyzed electrolysis) to reduce the organic load of
CW in parallel with hydrogen production. Overall, the
combined process resulted in the reduction of COD of the
effluent, while producing hydrogen at a maximum yield of
94.2 L ­H2/ kgVS. Rai et al. [41], investigated the optimum
concentration of selected trace elements (­Ni+2, ­Fe+2, and
­Mg+2) for photo-biohydrogen production by Rhodopseu-
domonas BHU 01 strain from the nutrient-poo and VFAs
rich spent medium resulting from dark-fermentation of CW
wastewater by Enterobacter aerogenes MTCC 2822. The
study showed that CW could be adopted as the effective
fermentation feedstock employing the dark-fermenting E.
aerogenes MTCC 2822. Moreover, Alvarado-Cuevas et al.
[42] reported through their study that nitrogen sources have
impact on hydrogen production by Escherichia coli using
CW as substrate. The research revealed that CW is an inex-
pensive potential raw material for hydrogen production by
fermentative processes, considering its high content of lac-
tose and lactalbumin; however, this complex protein is not
accessible for E. coli, and thus additional approaches are
needed. Dębowski et al. [43] determined the possibility of
applying psychrophilic bacteria for hydrogen production in
whey biofermentation process. In this study the effective-
ness of biohydrogen production from whey powder using
different groups of psychrophilic anaerobic bacteria (Gam-
maproteobacteria class—Rahnella aquatilis (9 strains) and
Firmicutes phylum: Carnobacterium maltaromaticum,
Trichococcus collinsii and Clostridium algidixylanolyticum
which had been earlier isolated from underground water
(3.7–5 m) and demersal lake water (17 m)), was investi-
gated. Kargi et al. [44] investigated the bio-hydrogen
production from cheese whey powder (CWP) solution
focusing on comparison of thermophilic and mesophilic
dark fermentation. The study showed that the thermophilic
dark fermentation of CWP solution is a promising approach
for biohydrogen production provided that the production
rates and yields were improved by simultaneous removal of
products (such as VFAs), using more effective bacterial
culture and optimum operating conditions. Kargi andUzun-
car [45] conducted experiments with diluted CW solution
in order to produce hydrogen by electrohydrolysis using
photo-voltaic cells (PVC) as source of electricity. Effects of
initial COD concentration on the rate and yield of hydrogen
gas production were investigated using a completely mixed
and sealed reactor with aluminum electrodes. The use of
PVC panels as source of electricity was found to be an
effective method for hydrogen gas production by electro-
hydrolysis of CW. Further improvements in ­H2 gas produc-
tion might be accomplished by using sequential fermenta-
tion and electro-hydrolysis of high strength CW solution
and operating the system under more intensive sunlight.
Perna et al. [46] tested an upflow anaerobic packed bed
reactor configuration at different organic loading rates to
produce hydrogen using CW.. Results showed that this pro-
cess could be a feasible and low-cost alternative for real-
scale applications that should be further studied to improve
the efficiency of hydrogen production. Rosales-Colunga
et al. [47] studied the fermentative hydrogen production by
E. coli using CW. This was the first work on biohydrogen
production by a genetically engineered E. coli strain, using
CW as substrate. Azbar et al. [48] investigated continuous
anaerobic fermentation processes for ­H2 production from
CW, under thermophilic conditions and different HRT val-
ues. The experimental results demonstrated that lower HRT
values (e.g. 1 day) caused an increase in hydrogen produc-
tion rate. The hydrogen percentage in the headspace was
not affected by any operational parameters studied.
Alcohols Production
Apart from the production of methane and hydrogen, eth-
anol production from CW has been also studied by many
authors. Sansonetti et al. [49] studied the feasibility of
bioethanol generation by batch fermentation of ricotta
cheese whey (“Scotta”). This substrate was characterised
by lactose ion of lactose between 4.5 and 5.0% (w/w) and
lower protein content compared to the traditional CW.
Traditional CW as well as deproteinized whey were also
tested in comparison to “scotta”. The microorganism used
for the fermentation process was the yeast Kluyveromyces
marxianu and the experiments conducted under anaerobic
conditions. Based on the results, the scotta is an excel-
lent substrate for fermentation achieving complete lactose
consumption in 13 h and great ethanol yield (97% of the
13

theoretical value). Joshi et al. [50] also studied the produc-
tion of bioethanol from CW fermentation using Kluyvero-
myces marxianus biofilm. The study showed, that fuel etha-
nol production by fermentation of CW (lactose) can be a
sustainable route for its valorization, solving the environ-
mental pollution problems caused due to its high BOD and
COD values. Another study using Kluyveromycess marxi-
anus for fermentative production of bioethanol from CW
was performed by Hadiyanto et al. [51] who examined the
effect of various temperatures (30, 35, 40 °C) to the ethanol
yields through fed-batch fermentation experiments. Results
showed that the highest biomass and ethanol concentrations
were achieved at 30 °C, with μ (0.186/h), Yp/s (0.21 g/g),
and Yx/s (0.32 g/g). Moreover, Diniz et al. [52] studied
the production of ethanol from CWP using Kluyveromy-
ces marxianus UFV-3. The results indicated that using K.
marxianus UFV-3 to convert lactose from CWP into etha-
nol is promising since yields close to the theoretical values,
were achieved, at wide range of temperatures, pH values,
and lactose concentrations, all of which are considered
crucial to the economic feasibility of using the perme-
ate as a raw material for ethanol production. Akbas et al.
[53] investigated the improvement of ethanol production by
engineered ethanologenic E. coli to express the hemoglobin
from the bacterium Vitreoscilla (VHb) providing evidences
that “VHb technology” may be useful for improving the
production of ethanol from waste and byproducts of vari-
ous sources. Kadar et al. [54] implemented a method for
bioethanol production from rye and wheat grain by uti-
lizing the inherent amylase activity from germination of
the seed. Simultaneous saccharification and fermentation
(SSF) was performed to reduce end-product inhibition of
the amylases during ethanol fermentation. The results of
the study showed that using organic CW as a cheap nutrient
source, the use of water and chemicals is minimized aiming
at reduced process costs. Another study was performed by
Guedes et al. [55] who evaluated the feasibility of bioetha-
nol production from Azorean whey permeate. The study
revealed that from a dairy plant that rejected 36,000,000 L
of whey, it is possible to obtain 620,000 L of bioethanol
which represents a financial advantage to Azorean econ-
omy and particularly for the dairy industry. Ozmihci and
Kargi [56] investigated the production of ethanol from
CW powder solution in a packed column bioreactor at dif-
ferent HRTs. The PCBR was found to be a compact and
effective reactor for ethanol production from CWP solution
with high ethanol yields as compared to the continuous sus-
pended cell bioreactors. Dragone et al. [57] examined the
optimal fermentation conditions for maximizing the etha-
nol production by Kluyveromyces fragilis from CWP. The
initial lactose concentration in deproteinized CWP solu-
tion exerted great influence on ethanol production by K.
fragilis, being the maximum product formation (80.95 kg/
13
Waste Biomass Valor
­ 3) obtained when using an initial lactose concentration
m
of 200 kg/m3. This value is about 4 times higher than that
achieved during the fermentation of non-concentrated CW
and represents thus an interesting alternative to decrease
the distillation costs for ethanol production. Silveira et al.
[58] studied the production of ethanol from CWP by K.
marxianus UFV-3 focusing on a flux analysis of oxido-
reductive metabolism as a function of lactose concentra-
tion and oxygen levels. In particular, the study focused on
the lactose concentration and the oxygen level that would
maximize ethanol yield. Raganati et al. [59] studied the
feasibility of bio-butanol production by continuous by
bioconversion of CW in a continuous packed bed reactor.
The CW conversion process was characterized in terms of
metabolites production (butanol included), lactose conver-
sion and biofilm mass. The study showed that under opti-
mized conditions, the performances were: butanol produc-
tivity 2.66 g/L/h, butanol concentration 4.93 g/L, butanol
yield 0.26 g/g, butanol selectivity of the overall solvents
production 82 wt%.
Electricity Production Through Microbial Fuel Cells
Apart from the above Tremouli et al. [60] studied the opera-
tion and characterization of a microbial fuel cell (MFC) fed
with pretreated (filter sterilized) CW at different organic
loads (0.35, 0.7, 1.5, 2.7 and 6.7 g COD/L respectively)
using n a two-chamber MFC. The best performance of the
cell was observed at the highest concentration of the CW
(6.7 g COD/L) corresponding to a maximum power den-
sity of approximately 46 mW/m2 Antonopoulou et al. [61]
investigated the possibility of electricity generation from
diluted cheese whey in a two-chamber mediator-less MFC
without any pretreatment.. In that study was shown that
power generation is possible by using diluted cheese whey
wastewater as energy source with a maximum power den-
sity obtained of 18.4 mW/m2 (normalized to the geomet-
ric area of the anodic electrode, which was 13.8 cm2), cor-
responding to a current density of 80 mA/m2 and a MFC
voltage of 0.23 V.. The results showed that pretreatment of
CW was necessary in order to minimize the competitive
activity of indigenous microbial cultures contained in CW
and to increase MFC performance [61].
Single Cell Protein Production
Ghaly et al. [62] studied a mathematical model which
described the kinetics of continuous production of single
cell protein (SCP) from CW using Kluyveromyces fragilis
which was developed from the basic principles of mass bal-
ances. The model takes into account the substrate utiliza-
tion for growth and maintenance and the effect of substrate
concentration and cell death rate on the net cell growth
Waste Biomass Valor
and substrate utilization during the fermentation process.
Results showed that the viable cell mass and lactose con-
sumption ranged from 1.3 to 34.3 g/L and from 74.31 to
99.02%, respectively.
Economic Aspects
Despite the relative high investment costs, AD is consid-
ered to be one of the most promising and economically
viable processes for treatment and valorization of various
wastes and wastewaters. The payback period ranges from 5
to 16 years, for operation under optimum or not optimized
conditions, respectively, a period that could be signifi-
cantly decreased through government financial incentives
[63]. Based on the above, AD could be a viable solution
for the valorization of wastewaters coming from Greek
cheese whey factories. For example, Diamantis et al. [9]
reported that for a small-to-medium size cheese produc-
tion plant, the application of a two-stage anaerobic diges-
tion process consisting of an acidogenic CSTR followed
by a UASB reactor, could result in a net profit of 0.94 €/
m3 CW. In this line, Spachos and Stamatis [64] conducted
an exergetic and economic analysis of an anaerobic treat-
ment system of whey, installed in one of the biggest dairy
industries in Greece (producing 150 m3/day of whey). The
produced biogas was subsequently burned and the energy
released was used for steam production for covering the
factory thermal needs. The Net Present Value (NPV) was
calculated based on the assumption that the whole capital
cost of the investment would be covered by a bank loan
received by the company. The results of the economic anal-
ysis revealed that the anaerobic treatment of whey is a prof-
itable and sustainable investment for the company, although
the exergetic efficiency of the system is rather low. Similar
results were also obtained by Chatzipaschali and Stamatis
[63] who performed an exergetic and economic analysis for
the same anaerobic treatment system for which however the
existing steam production was considered to be used for
covering both thermal and electrical needs of the factory. It
was shown by the economic analysis that this investment is
profitable for the company, with high savings and incomes
even from the first year of its operation. Another economi-
cally profitable solution for the factory could be the direct
conversion of the produced biogas to electricity solely, due
to the current high selling price of electricity to the net-
work. In this case, the electricity production is calculated
approximately to 1.5 MW. In all cases though, the scale is a
significant factor when evaluating the economic feasibility
of AD systems [65].
Regarding the other proposed processes for CW valori-
sation, it seems that there are a lot of limitations and bar-
riers to be overcome in order for the proposed technolo-
gies to be applicable in practice. Based on existing studies
regarding bioethanol production using CW, it has been
shown that an initial lactose concentration of 200 g/L in the
whey permeate, leads to the maximum ethanol yield pro-
viding the economic viability of the process [66]. However,
the supply chain of CW for bioethanol production is not
well defined, making effectiveness and implementation of
ethanol production from this substrate limited.
Conclusions
In conclusion, on the basis of the evaluation of data, the
prevailing CW valorisation practice by the Greek manu-
facturing units is its provision as feed for pigs. Apart from
CW, wastewater is also generated, which is mainly treated
in wastewater treatment plants in the case of medium scale
cheese manufacturing units. The most commonly applied
method was the extended aeration activated sludge pro-
cess. Cheese manufacturing units of smaller capacities
either dispose wastewater directly in the sewerage system
or they store it until it is transported for further treatment.
In international level, CW is currently highly utilised either
for the industrial production of other kind of cheese (e.g.
white cheese) or for the commercial manufacturing of other
whey derived products (e.g. sweet and acid whey powder,
reduced lactose whey, demineralized whey, whey pro-
tein concentrate, whey protein isolate). Apart from those
well-established practices, CW can be processed for the
production of valuable products and compounds that can
be valorised in the food and pharmaceutical industry; and
energy. So far, the anaerobic digestion process that leads
to the production of biogas has been extensively studied in
scientific publications and applied in large treatment plants.
Excepting methane production, CW has been studied as a
promising feedstock for fermentative hydrogen, ethanol and
butanol production, as well as for the production of elec-
tricity through MFC. Those alternatives need to be further
studied in terms of process feasibility in order to be applied
in industrial scale. However, whey products industrial pro-
duction, whey derived products manufacturing and applica-
tion of anaerobic digestion for CW utilisation can be con-
sidered as valorisation opportunities which could potential
add valued to the Greek cheese manufacturing sector.
Acknowledgements The authors would like to thank the European
Social Fund and the Hellenic Ministry of Education and Religious
Affairs, Cultures and Sports (Managing Authority) for funding the
project: FOODINBIO/2915 entitled “Development of an innovative,
compact system that combines biological treatment technologies for
the sustainable and environmental management of organic waste
streams that are produced from different types of food processing
industries”, in the framework of the Operational Programme Educa-
tional and Lifelong Learning (NSRF 2007–2013).
13
 Waste Biomass Valor

References Techno-economic investigation of water recovery and whey

powder production from whey using UF/RO and FO/RO inte-
grated membrane systems. Desalination. Water. Treat. 52, 123–
1. Valta, K., Kosanovic, T., Malamis, D., Moustakas, K., Loizidou,
133 (2014)
M.: Overview of water usage and wastewater management in
18. Prazeres, A.R., Carvalho, F., Rivas, J.: Fenton-like application
the food and beverage industry, Desalination. Water. Treat. 53
to pretreated cheese whey wastewater. J. Environ. Manage. 129,
(2015) 3335–3347, doi:10.1080/19443994.2014.934100.
199–205 (2013)
2. FEIR (Foundation for Economic and Industrial Research) Study:
19. Mollea, C., Marmo, L., Bosco, F.: Valorisation of cheese whey,
Facts and Figures (2014)
a by-product from the dairy industry, Chap. 24, In: Food Indus-
3. Eurostat Statistics Explained: Milk and milk product statistics.
try by Innocenzo Muzzalupo, doi:10.5772/53159, Available at:
Available at: http://ec.europa.eu/eurostat/statistics-explained/
http://www.intechopen.com/books/food-industry/valorisation-of-
index.php/Milk_and_milk_product_statistics (2016) Accessed
cheese-whey-a-by-product-from-the-dairy-industry (2013)
March 2016.
20. Prazeres, A.R., Carvalho, F., Rivas, J.: Cheese whey manage-
4. European Daily Association: Economic report 2016/17 Avail-
ment: a review. J. Environ. Manage. 110, 48–68 (2012)
able at: http://eda.euromilk.org/fileadmin/user_upload/Pub-
21. U.S. Dairy Export Council, : Reference Manual for U.S. Whey
lic_Documents/Facts_and_Figures/EDA_EWPA_Economic_
and Lactose Products, Available at: http://usdec.files.cms-plus.
Report_2016.pdf (2016) Accessed March 2016.
com/PDFs/2008ReferenceManuals/Whey_Lactose_Reference_
5. Commission Decision No 97/80/EC of 18 December 1996 lay-
Manual_Complete2_Optimized.pdf
ing down provisions for the implementation of Council Directive
22. Palatnik, D.R., Ostermann Porcel, M.V., González, U., Zaritzky,
96/16/EC on statistical surveys of milk and milk products (Text
N., Campdérros, M.E.: Recovery of caprine whey protein and
with EEA relevance), OJ L 24, 25.1.1997, pp. 26–49
its application in a food protein formulation., LWT – Food. Sci.
6. Valta, K., Damala, P., Orli, E., Papadaskalopoulou, C., Mousta-
Technol., doi:10.1016/j.lwt.2015.03.027 (2015)
kas, K., Malamis, D., Loizidou, M.: Valorisation opportunities in
23. Sanmartín, B., Díaz, O., Rodríguez-Turienzo, L., Cobos, A.:
the Greek slaughtering industry: current status and future poten-
Functional properties of caprine whey protein concentrates
tials. Waste. Biomass. Valor. (2015) 6:927–945, doi:10.1007/
obtained from clarified cheese whey Small. Rumin. Res. 110,
s12649-015-9368-1
52–56 (2013)
7. Valta, K., Aggeli, E., Papadaskalopoulou, C., Panaretou, V., Soti-
24. Galanakis, C.M., Chasiotis, S., Botsaris, G., Gekas, V.: Separa-
ropoulos, A., Malamis, D., Moustakas, K., Haralambous, K.-J.:
tion and recovery of proteins and sugars from Halloumi cheese
Adding value to olive oil production through waste and waste-
whey. Food Res. Int. 65, 477–483 (2014)
water treatment and valorisation: the case of Greece. Waste. Bio-
25. Folmer Corrêa, A.P., Joner Daroit, D., Fontoura, R., Meister
mass. Valor. (2015) 6:913–925, doi:10.1007/s12649-015-9373-4
Meira, S.M., Segalin, J., Brandelli, A.: Hydrolysates of sheep
8. Saddoud, A., Hassairi, I., Sayadi, S.: Anaerobic membrane
cheese whey as a source of bioactive peptides with antioxidant
reactor with phase separation for the treatment of cheese whey,
and angiotensin-converting enzyme inhibitory activities. Pep-
Bioresour. Technol. 98(11), 2102–2108 (2007)
tides. 61, 48–55 (2014)
9. Diamantis, V. I., Kapagiannidis, A. G., Ntougias, S., Tataki, V.,
26. Kumari Garsa, A., Kumariya, R., Kumar, A., Lather, P., Kapila,
Melidis, P., Aivasidis, A.: Two-stage CSTR–UASB digestion
S., Sood, S.K.: Industrial cheese whey utilization for enhanced
enables superior and alkali addition-free cheese whey treatment.
production of purified pediocin PA-1. LWT—Food. Sci. Tech-
Biochem. Eng. J. 84, 45–52, (2014)
nol.. 59, 656–665 (2014)
10. Gannoun, H., Khelifi, E., Bouallagui, H., Touhami, Y., Hamdi,
27. Watanabe, T., Shinozaki, Y., Suzuki, K., Koitabashi, M.,
M.: Ecological clarification of cheese whey prior to anaero-
Yoshida, S., Sameshima-Yamashita, Y., Kitamoto, H.K.: Produc-
bic digestion in upflow anaerobic filter. Bioresour. Technol. 99,
tion of a biodegradable plastic-degrading enzyme from cheese
6105–6111 (2008)
whey by the phyllosphere yeast Pseudozyma antarctica GB-4(1)
11. Rico, C., Muñoz, N., Fernández, J., Rico, J. L.: High-load anaer-
W J. Biosci. Bioeng. 118(2), 183–187 (2014)
obic co-digestion of cheese whey and liquid fraction of dairy
28. Galvão, C.M.A., Pinto, G.A., Jesus, C.D.F., Giordano, R.C.,
manure in a one-stage UASB process: limits in co-substrates
Giordano, R.L.C: Producing a phenylalanine-free pool of pep-
ratio and organic loading rate. Chem. Eng. J.. 262, 794–802
tides after tailored enzymatic hydrolyses of cheese whey. J. Food
(2015)
Eng. 91, 109–117 (2009)
12. Bertin, L., Grilli, S., Spagni, A., Fava, F.: Innovative two-stage
29. Sugai, M., Takakuwa, N., Ohnishi, M., Arai, I., Urashima, T.,
anaerobic process for effective codigestion of cheese whey and
Oda, Y.: Selection of lactic yeast producing glucosylceramide
cattle manure. Bioresour. Technol. 128, 779–783 (2013)
from cheese whey. Bioresour. Technol. 98, 3643–3646 (2007)
13. Comino, E., Riggio, V.A., Rosso, M.: Biogas production by
30. Dimitrellou, D., Kandylis, P., Sidira, M., Koutinas, A.A., Kourk-
anaerobic co-digestion of cattle slurry and cheese whey. Biore-
outas, Y.: Free and immobilized Lactobacillus casei ATCC 393
sour. Technol. 114, 46–53 (2012)
on whey protein as starter cultures for probiotic Feta-type cheese
14. Kavacik, B., Topaloglu, B.: Biogas production from co-digestion
production. J. Dairy. Sci. 97, 4675–4685 (2014)
of a mixture of cheese whey and dairy manure. Biomass and bio-
31. Di Cagno, R., De Pasquale, I., De Angelis, M., Buchin, S., Riz-
energy. 34(1321-1 3 2), 9 (2010)
zello, C.G., Gobbetti, M: Use of microparticulated whey protein
15. Dareioti, M. A., Kornaros, M.: Anaerobic mesophilic co-diges-
concentrate, exopolysaccharide-producing Streptococcus ther-
tion of ensiled sorghum, cheese whey and liquid cow manure
mophilus, and adjunct cultures for making low-fat Italian Caci-
in a two-stage CSTR system: effect of hydraulic retention time.
otta-type cheese. J. Dairy Sci. 97, 72–84 (2014)
Bioresour. Technol. 175, 553–562 (2015)
32. Reyes, J. C., Celis, L. B., Mondragon, F. A., Flores, E. R.:
16. Frigon, J.-C., Breton, J., Bruneau, T., Moletta, R., Guiot, S.R.:
Decreasing methane production in hydrogenogenic UASB reac-
The treatment of cheese whey wastewater by sequential anaero-
tors fed with cheese whey. Biomass. Bioenergy. 63, 101–108
bic and aerobic steps in a single digester at pilot scale. Bioresour.
(2014)
Technol. 100, 4156–4163 (2009)
33. Gomez-Romero, J., Gonzalez-Garcia, A., Chairez, I., Torres, L.,
17. Aydiner, C., Sen, U., Topcu, S., Sesli, D., Ekinci, D., Altınay,
Garcıa-Pena, E.I.: Selective adaptation of an anaerobic microbial
A.D., Ozbey, B., Koseoglu-Imer, D.U., Keskinler, B.:
community: Biohydrogen production by codigestion of cheese

13
Waste Biomass Valor
whey and vegetables fruit waste. Int. J. Hydrog. Energy. 39,
12541–12550 (2014)
34. Davila-Vazquez, G., Cota-Navarro, C.B., Rosales-Colunga,
L.M., Leon-Rodrıguez, A., Razo-Flores, E.: Continuous biohy-
drogen production using cheese whey: Improving the hydrogen
production rate. Int. J. Hydrog. Energy. 34, 4296–4304 (2009)
35. Castelló, E., García y Santos, C., Iglesias, T., Paolino, G., Wen-
zel, J., Borzacconi, L., Etchebehere, C.: Feasibility of biohydro-
gen production from cheese whey using a UASB reactor: links
between microbial community and reactor performance. Int. J.
Hydrog. Energy. 34, 5674–5682 (2009)
36. Venetsaneas, N., Antonopoulou, G., Stamatelatou, K., Kornaros,
M., Lyberatos, G.: Using cheese whey for hydrogen and meth-
ane generation in a two-stage continuous process with alternative
pH controlling approaches. Bioresour. Technol. 100, 3713–3717
(2009)
37. Antonopoulou, G., Stamatelatou, K., Venetsaneas, N., Kornaros,
M., Lyberatos, G.: Biohydrogen and methane production from
cheese whey in a two –stage anaerobic process. Ind. Eng. Chem.
Res., 47(15), 5227–5233, (2008)
38. Fernández, C., Carracedo, B., Martínez, E.J., Gómez, X., Morán,
A.: Application of a packed bed reactor for the production of
hydrogen from cheese whey permeate: effect of organic loading
rate. J. Environ. Sci. Health,. 49, 210–217 (2014)
39. Seo, Y.H., Yun, Y.M., Lee, H., Han, J.-I.: Pretreatment of cheese
whey for hydrogen production using a simple hydrodynamic
cavitation system under alkaline condition. Fuel. 150, 202–207
(2015)
40. Moreno, R., Escapa, A., Cara, J., Carracedo, B., Gómez, X.: A
two-stage process for hydrogen production from cheese whey:
Integration of dark fermentation and biocatalyzed electrolysis.
Int. J. Hydrog. Energy. 40, 168–175 (2015)
41. Rai, P.K., Asthana, R.K., Singh, S.P.: Optimization of photo-
hydrogen production based on cheese whey spent medium. Int. J.
Hydrog. Energy. 39, 7597–7603 (2014)
42. Alvarado-Cuevas, Z.D., Ordoñez Acevedo, L.G., Ornelas Salas,
J.T., De León-Rodríguez, A.: Nitrogen sources impact hydrogen
production by Escherichia coli using cheese whey as substrate
New Biotechnol. 30(6), 585–590 (2013)
43. Dębowski, M., Korzeniewska, E., Filipkowska, Z., Zieliński,
M., Kwiatkowski, R.: Possibility of hydrogen production dur-
ing cheese whey fermentation process by different strains of
psychrophilic bacteria, Int. J. Hydrog. Energy 39, 1972–1978,
(2014)
44. Kargi, F., Eren, N.S., Ozmihci, S.: Hydrogen gas production
from cheese whey powder (CWP) solution by thermophilic dark
fermentation. Int. J. Hydrog. Energy. 37, 2260–2266 (2012)
45. Kargi, F., Uzuncar, S.: Electro-hydrolysis of cheese whey solu-
tion for hydrogen gas production and chemical oxygen demand
(COD) removal using photo-voltaic cells (PVC). Int. J. Hydrog.
Energy. 37, 15841–15849 (2012)
46. Perna, V., Castello, E., Wenzel, J., Zampol, C., Fontes Lima,
D.M., Borzacconi, L., Varesche, M.B., Zaiat, M., Etchebehere,
C.: Hydrogen production in an upflow anaerobic packed bed
reactor used to treat cheese whey. Int. J. Hydrog. Energy. 38,
54–62 (2013)
47. Rosales-Colunga, L.M., Razo-Flores, E., Ordonez, L.G., Ala-
triste-Mondragon, F., De Leon-Rodrıguez, A: Hydrogen produc-
tion by Escherichia coli DhycA DlacI using cheese whey as sub-
strate. Int. J. Hydrog. Energy. 35, 491–499 (2010)
48. Azbar, N., Dokgöz, F.T.Ç., Keskin, T., Korkmaz, K.S., Syed,
H.M.: Continuous fermentative hydrogen production from
cheese whey wastewater under thermophilic anaerobic condi-
tions. Int. J. Hydrog. Energy. 34, 7441–7447 (2009)
49. Sansonetti, S., Curcio, S., Calabro, V., Iorio, G.: Bio-ethanol pro-
duction by fermentation of ricotta cheese whey as an effective
alternative non-vegetable source. Biomass. Bioenerg. 33, 1687–
1692 (2009)
50. Joshi, Y., Poletto, M., Senatore, B.: Bioethanol from cheese
whey fermentation using Kluyveromyces marxianus biofilm. J.
Biotechnol. 11, 178–179 (2010)
51. Hadiyanto, Ariyanti, D., Aini, A.P., Pinundi, D.S.: Optimization
of ethanol production from whey through fed-batch fermentation
using Kluyveromyces marxianus Energy Procedia 47, 108–112
(2014)
52. Diniz, R.H.S., Marina Q.R.B. Rodrigues, Fietto, L.G., Passos,
F.M.L., Silveira, W.B.: Optimizing and validating the produc-
tion of ethanol from cheese whey permeate by Kluyveromyces
marxianus UFV-3, Biocatalysis and Agricultural. Biotechnology.
3, 111–117 (2014)
53. Akbas, M.Y., Sar, T., Ozcelik, B.: Improved ethanol production
from cheese whey, whey powder, and sugar beet molasses by
“Vitreoscilla hemoglobin expressing” Escherichia coli, Biosci,
Biotechnol, Biochem, 78(4), 687–694 (2014)
54. Kádár, Z., Christensen, A.D., Hedegaard Thomsen, M., Bjerre,
A.-B.: Bioethanol production by inherent enzymes from rye and
wheat with addition of organic farming cheese whey. Fuel. 90,
3323–3329 (2011)
55. Guedes, C., Garcia, D., Lima, E., Baptista, J.: Hydrolysis of lac-
tose from bovine whey for subsequent fermentation to bio-etha-
nol. J. Biotechnol. 11, 139–139 (2010)
56. Ozmihci, S., Kargi, F.: Ethanol production from cheese whey
powder solution in a packed column bioreactor at different
hydraulic residence times Biochem. Eng. J. 42, 180–185 (2008)
57. Dragone, G., Mussatto, S.I., Almeida e Silva, J.B., Teixeira, J.A.:
Optimal fermentation conditions for maximizing the ethanol pro-
duction by Kluyveromyces fragilis from cheese whey powder.
Biomass Bioenergy. 35, 1977–1982 (2011)
58. Silveira, W.B., Passos, F.J.V., Mantovani, H.C., Passos, F.M.L:
Ethanol production from cheese whey permeate by Kluyveromy-
ces marxianus UFV-3: a flux analysis of oxido-reductive metab-
olism as a function of lactose concentration and oxygen levels.
Enzyme Microb. Technol. 36, 930–936 (2005)
59. Raganati, F., Olivieri, G., Procentese, A., Russo, M.E., Sala-
tino, P., Marzocchella, A.: Butanol production by bioconversion
of cheese whey in a continuous packed bed reactor. Bioresour.
Technol. 138, 259–265 (2013)
60. Tremouli, A., Antonopoulou, G., Bebelis, S., Lyberatos, G.:
Operation and characterization of a microbial fuel cell fed with
pretreated cheese whey at different organic loads. Bioresour.
Technol. 131, 380–389 (2013)
61. Antonopoulou, G., Stamatelatou, K., Bebelis, S., Lyberatos, G.:
Electricity generation from synthetic substrates and cheese whey
using a two chamber microbial fuel cell Biochem. Eng. J. 50,
10–15 (2010)
62. Ghaly, A.E., Kamal, M., Correia, L.R.: Kinetic modelling of
continuous submerged fermentation of cheese whey for sin-
gle cell protein production. Bioresour. Technol. 96, 1143–1152
(2005)
63. Chatzipaschali, A.A., Stamatis, A.G.: Biotechnological utiliza-
tion with a focus on anaerobic treatment of cheese whey: current
status and prospects. Energies 5, 3492–3525 (2012)
64. Spachos, T., Stamatis, A.: Thermal analysis and optimization
of an anaerobic treatment system of whey. Renew. Energy. 36,
2097–2105 (2011)
65. Greer, D.: Anaerobic digestion for smaller dairies. BioCycle. 51,
24–26 (2010)
66. Das, M., Raychaudhuri, A., Ghosh, S.K.: Supply Chain of
bioethanol production from whey: a Review. Procedia Environ.
Sci. 35, 833–846 (2016)
13