Process Biochemistry 40 (2005) 2583–2595

www.elsevier.com/locate/procbio

Anaerobic treatment of dairy wastewaters: a review

Burak Demirel, Orhan Yenigun *, Turgut T. Onay
Institute of Environmental Sciences, Bogazici University, Bebek, 34342 Istanbul, Turkey
Received 11 May 2004; accepted 3 December 2004

Abstract

Anaerobic treatment is often reported to be an effective method for treating dairy effluents. The objective of this paper is to summarize
recent research efforts and case studies in anaerobic treatment of dairy wastewaters. The main characteristics of industrial dairy waste streams
are identified and the anaerobic degradation mechanisms of the primary constituents in dairy wastewaters, namely carbohydrates (mainly
lactose), proteins and lipids are described. Primary attention is then focused on bench–pilot–full-scale anaerobic treatment efforts for dairy
waste effluents. Combined (anaerobic–aerobic) treatment methods are also discussed. Finally, areas where further research and attention are
required are identified.
# 2005 Elsevier Ltd. All rights reserved.

Keywords: Anaerobic treatment; Dairy wastewaters; Acidogenesis; Lipids degradation; Proteins degradation

1. Introduction malfunctions or operational errors [4,6]. Dairy wastewaters

The dairy industry, like most other agro-industries,
generates strong wastewaters characterized by high biolo-
gical oxygen demand (BOD) and chemical oxygen demand
(COD) concentrations representing their high organic
content [1]. Furthermore, the dairy industry is one of the
largest sources of industrial effluents in Europe. A typical
European dairy generates approximately 500 m3 of waste
effluent daily [2]. Dairy waste effluents are concentrated in
nature, and the main contributors of organic load to these
effluents are carbohydrates, proteins and fats originating
from the milk [3,4]. Since dairy waste streams contain high
concentrations of organic matter, these effluents may cause
serious problems, in terms of organic load on the local
municipal sewage treatment systems [3]. In addition to
environmental problems that can result from discharge of
dairy wastewaters, introduction of products such as milk
solids into waste streams also represents a loss of valuable
product for the dairy facilities [5]. Most of the wastewater
volume generated in the dairy industry results from cleaning
of transport lines and equipment between production cycles,
cleaning of tank trucks, washing of milk silos and equipment
* Corresponding author. Tel.: +90 212 3596946; fax: +90 212 2575033.
E-mail address: yeniguno@boun.edu.tr (O. Yenigun).
are treated using physico-chemical and biological treatment
methods. However, since the reagent costs are high and the
soluble COD removal is poor in physical–chemical
treatment processes, biological processes are usually
preferred [7]. Among biological treatment processes,
treatment in ponds, activated sludge plants and anaerobic
treatment are commonly employed for dairy wastewater
treatment [8]. In contrast the contrary, high energy
requirements of aerobic treatment plants is a significant
drawback of these processes. COD concentrations of dairy
effluents vary significantly; moreover, dairy effluents are
warm and strong, enabling them ideal for anaerobic
treatment [2]. Furthermore, no requirement for aeration,
low amount of excess sludge production and low area
demand are additional advantages of anaerobic treatment
processes, in comparison to aerobic processes. The aim of
this paper is to summarize the recent research efforts and
case studies in anaerobic treatment of dairy waste effluents.
In the paper, the general characteristics of dairy waste
streams are identified and the anaerobic degradation
mechanisms of the main constituents of dairy wastewaters,
namely carbohydrates, proteins and lipids, are explained.
Anaerobic treatment practices of dairy wastewaters, as
bench, pilot and full-scale efforts, are subsequently
introduced overall in detail. Combined (anaerobic–aerobic)

0032-9592/$ – see front matter # 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.procbio.2004.12.015
2584 B. Demirel et al. / Process Biochemistry 40 (2005) 2583–2595

treatment systems for dairy wastewaters are also summar-
ized briefly. Finally, areas where particular research and
more attention required in the near future are identified.
2. General characteristics of dairy wastewaters
Wastewaters from the dairy industry are usually
generated in an intermittent way, so the flow rates of these
effluents change significantly. High seasonal variations are
also encountered frequently and correlate with the volume of
milk received for processing; which is typically high in
summer and low in winter months [9]. Moreover, since the
dairy industry produces different products, such as milk,
butter, yoghurt, ice-cream, various types of desserts and
cheese, the characteristics of these effluents also vary
greatly, depending on the type of system and the methods of
operation used [7]. The use of acid and alkaline cleaners and
sanitizers in the dairy industry additionally influences
wastewater characteristics and typically results in a highly
variable pH [4,6,10]. Actually, information about the general
characteristics of dairy wastewaters from full-scale opera-
tions in literature is scarce. Only one comprehensive study
has been encountered, which provides extensive information
about the particular characteristics of dairy wastewaters
from various full-scale operations [6]. A summary of data
obtained from literature for general properties of dairy waste
effluents from full-scale operations is given in Table 1 [11–
19]. High COD concentrations indicate that dairy industry
wastewaters are strong and fluctuating in nature. Significant
fractions of the organic components and nutrients in dairy
waste streams are derived from milk and milk products. In
industrial dairy wastewaters, nitrogen originates mainly
from milk proteins, and is present in various forms; either an
organic nitrogen (proteins, urea, nucleic acids), or as ions
such as NH4+, NO2 and NO3 . Phosphorus is found mainly
in inorganic forms; as orthophosphate (PO43 ) and poly-
phosphate (P2O74 ), as well as organic forms [20]. Concen-
trations of suspended solids (SS) and volatile suspended
solids (VSS) are also used to evaluate wastewater strength
and treatability [6]. Suspended solids in dairy wastewaters
originate from coagulated milk, cheese curd fines or
flavoring ingredients [21]. Concentrations of selected
elements, namely sodium (Na), potassium (K), calcium
(Ca), magnesium (Mg), iron (Fe), cobalt (Co), nickel (Ni)
and manganese (Mn), are also given in Table 2. Particularly
high Na concentrations point out the use of large amount of
alkaline cleaners at dairy plants. The concentrations of

Table 1
Characteristics of dairy waste effluents
Effluent COD BOD5 pH Alkalinity Suspended Volatile Total TKN Total Reference
type (mg/l) (mg/l) (units) (mg CaCO3/l) solids suspended solids (mg/l) phosphorus
(mg/l) solids (mg/l) (mg/l) (mg/l)
Creamery 2000–6000 1200–4000 8–11 150–300 350–1000 330–940 50–60 [4]
Not given 980–7500 680–4500 300 [9]
Mixed dairy 1150–9200 6–11 320–970 340–1730 255–830 2705–3715 14–272 8–68 [11]
processing
Cheese whey 68814a 1462a 379a [12]
Cheese 1000–7500 588–5000 5.5–9.5 500–2500 [13]
Fresh milk 4656a 6.92a [14]
Cheese 5340a 5.22a [14]
Milk powder/ 1908a 5.80a [14]
butter
Mixed dairy 63100a 3.35a 12500a 12100a 53000a [15]
processing
Cheese whey 61000a 1780a 1560a 980a 510a [16]
Cheese 4.7a 2500a 830a 280a [17]
Not given 4.4–9.4 90–450 [18]
Fluid milk 950–2400 500–1300 5.0–9.5 90–450 [19]
a
Mean concentrations are reported.

Table 2
Concentrations of selected elements in dairy wastewaters
Effluent type Na (mg/l) K (mg/l) Ca (mg/l) Mg (mg/l) Fe (mg/l) Co (mg/l) Ni (mg/l) Mn (mg/l) Reference
Creamery 170–200 35–40 35–40 5–8 2–5 0.05–0.15 0.5–1.0 0.02–0.10 [4]
Cheese/whey 735a 42.8a 47.7a 11.4a [6]
Cheese/alcohol 423a 41.2a 54.3a 8.3a [6]
Cheese/beverages 453a 8.6a 33.6a 16.9a [6]
Cheese/whey 419a 35.8a 52.3a 11.0a [6]
Mixed dairy 123–2324 8–160 12–120 2–97 0.5–6.7 0 0–0.13 0.03–0.43 [11]
Cheese 720–980 530–950 [13]
a
Mean concentrations are reported.
 B. Demirel et al. / Process Biochemistry 40 (2005) 2583–2595
heavy metals, such as copper (Cu), nickel (Ni) and zinc (Zn)
were reported to be in a range that would not affect adversely
the performance of a biological treatment step [6,11].
As stated above, dairy wastewater is composed of easily
degradable carbohydrates, mainly lactose, as well as less
biodegradable proteins and lipids [22]. In cheese-processing
wastewater, 97.7% of total COD was accounted for by lactose,
lactate, protein and fat [15]. Thus, dairy wastewater can easily
be defined as a complex type of substrate [22–24]. Lactose is
the main carbohydrate in dairy wastewater and is a readily
available substrate for anaerobic bacteria. Anaerobic metha-
nation of lactose needs a cooperative biological activity from
acidogens, acetogens and methanogens [25]. Anaerobic
fermentation of lactose yields organic acids, namely acetate,
propionate, iso- and normal-butyrate, iso- and normal
valerate, caproate, lactate, formate and ethanol [26,27].
Two possible carbon flow schemes were proposed for
acidogenic fermentation of lactose; carbon flow from
pyruvate to butyrate and lactate, both occurring in parallel
[28]. The presence of high carbohydrate concentrations in
synthetic dairy wastewater was found to reduce the amount of
proteolytic enzymes synthesized, resulting in low levels of
protein degradation [22]. It was previously reported that
carbohydrates could suppress the synthesis of exopeptidases,
a group of enzymes facilitating protein hydrolysis [29].
Anaerobic degradation of proteins and the effects of ammonia
on this mechanism were recently investigated in detail [30–
35]. Casein is the major protein in milk composition and in
dairy effluents. When fed to acclimated anaerobic reactors,
degradation of casein is very fast and the degradation products
are non-inhibitory [3].
Lipids are potentially inhibitory compounds, which can
always be encountered during anaerobic treatment of dairy
wastewaters. There is little information available in literature
about the anaerobic digestibility of lipids. During anaerobic
degradation, lipid is firstly hydrolyzed to glycerol and long
chain fatty acids (LCFAs), followed by b-oxidation,
producing acetate and hydrogen [29]. The biodegradation
of lipids is difficult due to their low bioavailability [36].
Glycerol, a compound formed as a result of lipid hydrolysis,
was found to be a non-inhibitory compound [3], while,
LCFAs were particularly reported to be inhibitory to
methanogenic bacteria [37]. The inhibitory effects of lipids
in anaerobic processes can mainly be correlated to the
presence of LCFAs, which cause retardation in methane
production [38]. Lipids do not cause serious problems in
aerobic processes, however, they sometimes affect conven-
tional single-phase anaerobic treatment processes adversely
[39,40]. Unsaturated LCFAs seemed to have a greater
inhibitory effect than saturated LCFAs. Unsaturated LCFAs
strongly inhibited methane production from acetate and
moderately inhibited b-oxidation. Thus, unsaturated LCFAs
should be saturated to prevent lipid inhibition in anaerobic
processes [40]. Difficulties experienced with the presence of
lipids in anaerobic treatment processes have been previously
reported in literature[41–44].
2585
3. Conventional (single-phase) anaerobic treatment
of dairy effluents
Anaerobic treatment processes are favourable methods
for treating dairy waste effluents, in comparison to aerobic
processes, due to their well-known benefits for treating
industrial wastewaters, particularly from agricultural indus-
tries with a high organic content [45–47]. Anaerobic
treatment applications for dairy industry wastewaters have
been evaluated in a number of previous studies [48–59].
More recent information about anaerobic treatment prac-
tices of dairy waste streams are also displayed in Table 3.
In treatment studies of dairy wastewaters, anaerobic filters
have recently been used. If the particular dairy effluents
contain low concentrations of suspended solids, then
anaerobic filter reactors are generally suitable for biological
treatment. A laboratory-scale plastic medium anaerobic filter
reactor provided average COD removal rates between 78 and
92%, at a hydraulic retention time (HRT) of 4 days [60,61]. In
addition, the start-up performance of the anaerobic filters
treating dairy wastewater, in terms of COD removal, was not
significantly affected by temperature variations between 21
and 30 8C [62]. The effects of porous and non-porous support
media in anaerobic filter reactors on thermophilic anaerobic
treatment of ice-cream wastes were investigated extensively
[63,64]. At high loading rates, anaerobic filter with porous
support media performed more satisfactorily [63]. The
performance of porous and non-porous media in an upflow
anaerobic filter (UAF) treating wastewaters from a milk
bottling factory was also investigated in a more recent work
[65]. The reactor with non-porous packing showed instability
above an organic loading rate (OLR) of 4 kg COD/(m3 day),
while the reactor with porous packing was still stable at OLRs
up to 21 kg COD/(m3 day). The results of a pilot trial showed
that the anaerobic filter reactor, treating wastewaters from an
ice-cream manufacturer, achieved a mean COD removal rate
of 70%, at an average load of 5.5 kg COD/(m3 day) [66]. Low
temperature kinetics of anaerobic filters treating dairy
wastewaters were determined for various HRT ranges, using
three reactors operated at 12.5, 21 and 30 8C [67]. A
relationship was developed between the temperature of the
system and the first-order rate constant for each anaerobic
filter reactor. In another work, an upflow anaerobic filter
reactor (UAF) was used to treat very dilute dairy wastewater,
in an OLR range of between 0.117 and 1.303 g volatile solids
(VS)/(l day) and at an HRT range between 18.8 and 2 days
[68]. At 5, 4 and 3 days of HRT, effluent SS and COD
concentrations satisfied the effluent discharge limits. A pilot-
scale upflow anaerobic filter reactor treating dairy waste-
waters provided more than 85% COD and 90% BOD removal,
at an OLR of 6 kg COD/(m3 day) [69]. At 20 h of HRT, the
percentage of methane in the biogas produced by the UFAF
was found to be in a range between 75 and 85%, with a
corresponding methane yield of about 0.33 m3 CH4/kg COD
removed. The system could generate approximately 770 l
methane (CH4)/day. A comparative study of staged and
 2586
Table 3
Anaerobic treatment performance levels for dairy wastewaters
Waste type Reactor type HRT Loading Temperature Methane yield Removal (%) Application Reference
(8C) (m3/kg CODra) status
Cheese whey Downflow fixed-film 4.9 (day) 13 (kg COD/(m3 day)) 0.28 75 (COD) Pilot scale [16]
Cheese whey Downflow fixed-film 6.6 (day) 8.3 (kg COD/(m3 day)) 0.34 76 (COD) Pilot scale [16]
Anaerobic filter 4 (day) 30–21–12.5 92–85–78 (COD) Laboratory scale [60,61]
Ice-cream Anaerobic filter 5.5 (kg COD/(m3 day)) 75 (COD) Pilot scale [66]
Anaerobic filter 6 (kg COD/(m3 day)) 0.32–0.34 85 (COD) Pilot scale [69]
Anaerobic filter 0.5 (day) Up to 21 (kg COD/(m3 day)) 80 (COD) [72]
Raw milk Anaerobic filter 5–6 (kg COD/(m3 day)) 90 (COD) Full scale [73]
Cheese whey UASB >97 (COD) Laboratory scale [75]

B. Demirel et al. / Process Biochemistry 40 (2005) 2583–2595

Cheese production UASB 31 (g COD/(l day)) 90 (COD) Laboratory scale [78]
wastewater
Whey permeate UASB 0.4–5 (day) 99–64.2 (COD) Laboratory scale [80]
Effluent of an Hybrid UASB 5 (h) 8.5 (g COD/(l day)) 30 87 (COD) Laboratory scale [19]
integrated plant
Hybrid UASB 1–8 (g COD/(l day)) 30 92 Laboratory scale [82]
Cheese production 6 2–7.3 85–99
wastewater
10–20 2.3–4.5 35 79–91 Laboratory scale [17]
UASB 30–40 (day) 1.5–1.9 (g COD/(l day)) 81
UASB 90 (COD) [83]
3
Synthetic dairy Hybrid 4.1–1.7 (day) 0.82–6.11 (kg COD/(m day)) 35 0.354 (at 1.7 90–97 (COD) Laboratory scale [85]
effluent days HRT)
Cheese whey Hybrid 2 (day) Up to 11 (kg COD/(m3 day)) >95 (COD) Laboratory scale [86]
Synthetic non-fat ASBR 6 (h) 5 62 (COD), 75 (BOD5) Laboratory scale [87]
dry milk
ASBR 3–6 (day) 2–4 (g VS/(l day)) 26–50 (VS) Laboratory scale [88,89]
Upflow packed-bed 5–14.29 (kg COD/(m3 day)) 93.8–98.5, 72.5–84 (COD) [92]
Washing and 90.4 (COD) Laboratory scale [93]
rinsing waters
Cheese whey Rotating biological 3 (day) 37 85 (COD) [95]
contact reactor
Upflow anaerobic solid 4.5 (h) 20 98 (lipid) Laboratory scale [96]
removal reactor
Cheese whey Multichamber bioreactor 2 (day) 37 83 (COD) [98]
Synthetic ice-cream CSTR 7.45–5.99–4.60 35 98–97–96–94–92 (SCOD)b Laboratory scale [100]
wastewater –3.76–2.99 (day)
a
COD removal.
b
Soluble COD.
 B. Demirel et al. / Process Biochemistry 40 (2005) 2583–2595
non-staged anaerobic filters treating synthetic dairy waste-
waters was conducted at an HRT of 2 days and in a substrate
concentration range of from 3 to 12 g COD/l [70]. The authors
reported that the performances of both reactors were very
similar under the operating conditions tested. Laboratory-
scale single-fed (SFR) and multi-fed (MFR) upflow anaerobic
filters treating cheese whey were operated at OLRs above
20 kg COD/(m3 day) [71]. It was observed that the feeding
regime affected both biomass concentration and activity. The
specific activities of the different trophic groups were found to
be higher in the MFR. An upflow anaerobic filter reactor
yielded an average of 80% COD removal in an OLR range up
to 21 kg COD/(m3 day) when treating dairy wastewater [72].
Recently, the performance of an industrial-scale anaerobic
filter treating raw milk discharged by quality control
laboratories was reported [73]. Higher than 90% of COD
removal could be attained, with an OLR maintained around 5–
6 kg COD/(m3 day). Moreover, the fat content in dairy
wastewater could successfully be degraded by the anaerobic
filter reactor.
Upflow anaerobic sludge blanket (UASB) reactors have
been successfully employed for dairy wastewater treatment
in full-scale applications for almost two decades [74].
Biological treatment of a cheese-producing wastewater by a
laboratory-scale UASB reactor was reported for influent
wastewater concentrations between 12 and 60 g COD/l [17].
COD removal rates varied between 85 and 99%, at an HRT
of 6 days and in an OLR range of from 2 to 7.3 g COD/
(l day), while removal rates were around 81% in an HRT
range between 30 and 40 days. Anaerobic treatability studies
of dairy effluents from a large integrated industry processing
milk were carried out using a laboratory-scale hybrid UASB
reactor [19]. At an OLR of 8.5 g COD/(l day) and an HRT of
5 h, 87% COD removal was achieved at 30 8C. Another
laboratory-scale investigation pointed out that more than
97% COD reduction could be achieved in an UASB reactor
during anaerobic treatment of cheese whey [75]. Methanol
addition during start-up of UASB reactors treating a dairy
waste from raw ice-cream production facility provided rapid
granulation of biomass and enhanced the settling velocity
and specific activity of the sludge [76]. However, methanol
addition also resulted in severe biomass wash-out from the
system. During laboratory-scale anaerobic digestion of
cheese whey, increased substrate loadings led to the failure
of the UASB reactor, in a range of influent substrate
concentration from 4.5 to 38.1 g COD/l, at an HRT of 5 days
[77]. Anaerobic treatment of cheese-production wastewater
using a laboratory-scale UASB reactor provided a COD
removal of about 90%, at an OLR of 31 g COD/(l day) [78].
Furthermore, OLR peaks above 45 g COD/(l day) yielded a
COD reduction between 70 and 80%. Sudden increase in the
OLR was accompanied by biomass granulation, resulting in
a more stable reactor operation during waste treatment. The
kinetic model and the kinetic coefficients of laboratory-scale
continuous UASB reactors treating whey permeate were
determined, in an HRT range between 0.4 and 5 days and at a
2587
constant influent substrate concentration of 10.4  0.2 g
COD/l [79]. The maximum substrate utilization rate (k), half
saturation coefficient (KL), yield coefficient (Y) and decay
rate coefficient (Kd) were determined to be 0.941,
0.773 kg COD/(kg VSS day), 0.153 kg VSS/(kg COD) and
0.022 day 1, respectively. Under the same HRT and influent
substrate concentration, COD removal efficiency in the
UASB reactor ranged between 64.2 and 99% [80]. The
anaerobic digestion of cheese whey was investigated, in
terms of instability caused by the strength of the influent in
an UASB reactor [81]. For proper system stability, the
optimum influent substrate concentration was determined to
be between 25 and 30 g COD/l at an HRT of 5 days. A
laboratory-scale combined system designed by converting
the flow mixing chamber of an anaerobic filter into an UASB
resulted in an organic matter removal of 92% for dairy
wastewater, in an OLR range of between 1 and 8 g COD/
(l day), at 30 8C [82]. Dairy wastewaters containing high
concentrations of fat and grease were treated by an UASB
reactor [83]. COD removal was reported to be about 90%.
Furthermore, a new generation of more advanced anaerobic
reactor systems have also been developed, based on the
UASB system. A successful version of this concept is the
internal circulation (IC) reactor [84]. The IC reactor system
is able to handle high upflow liquid and gas velocities, which
enables treatment of low strength wastes at short HRTs, as
well as treatment of high-strength effluents at very high
volumetric loading rates feasible. Recently, feasibility of
using UASB reactors for dairy wastewater treatment was
explored by operating two types of UASB reactors [85]. The
reactors were operated at an HRT range between 3 and 12 h,
and on loadings ranging from 2.4 to 13.5 kg COD/(m3 day).
At 3 h, maximum COD reduction ranged between 95.6 and
96.3%, while at 12 HRT reductions were around 92–90%,
for both reactors.
In addition to anaerobic filters and UASB reactors, hybrid
digesters and anaerobic sequencing batch reactors (ASBR)
are also employed for treating dairy effluents. A mesophilic
laboratory-scale hybrid anaerobic digester, combining an
upflow sludge blanket and a fixed-bed design, was used to
treat synthetic dairy effluent with an influent substrate
concentration of 10 g COD/l [86]. At an HRT range between
4.1 and 1.7 days, COD removal rates between 90 and 97%
could be achieved, at an OLR range between 0.82 and
6.11 kg COD/(m3 day). The anaerobic digester provided a
methane yield of 0.354 m3 CH4/kg COD removed at an HRT
of 1.7 days. Anaerobic treatment of a high-strength acidic
cheese whey by a laboratory-scale upflow hybrid reactor
resulted in removal efficiencies of more than 95%, at 2 days
of HRT and up to an OLR of about 11 kg COD/(m3 day)
[87]. ASBRs also provide high treatment efficiencies for
dairy effluents. The laboratory-scale ASBR system was
reported to provide soluble COD and BOD5 removal rates of
62 and 75%, respectively, at an HRT of 6 h, at 5 8C, for a
synthetic substrate of non-fat dry milk [88]. In a temperature
range between 5 and 20 8C, and at an HRT range between 24
2588 B. Demirel et al. / Process Biochemistry 40 (2005) 2583–2595
and 6 h, soluble organic removal rates ranged between 62
and 90% for COD, and 75 and 90% for BOD5. In another
laboratory-scale work, two-stage thermophilic ASBR
systems provided volatile solids removal of 26–44%, while
mesophilic ASBR systems achieved VS removal between 26
and 50% for dairy wastewater [89,90]. The systems were
operated in an OLR range of 2–4 g VS/(l day) and at HRTs
of 3 and 6 days. The purification performance and the basic
fundamentals for the design of an ASBR used for treating
concentrated dairy wastewater were investigated in a more
recent study [91]. The maximum loading rate was
determined to be 6 g COD/(l day) for stable operation,
since higher loadings were reported to cause problems, such
as sludge removal and purification efficiency declines.
Actually, more studies focusing on different aspects of
anaerobic treatment of dairy wastewaters were performed
during the last decade. The changes in the microbial ecology
in four different pilot-scale digesters, namely anaerobic
contact, anaerobic filter, anaerobic expanded/fluidized bed
reactor and UASB reactor, all treating ice-cream wastewater,
were comprehensively examined during start-up [92]. The
authors reported that the reactor configuration did not play
an important role in causing changes in the microbial
community. A two-stage upflow packed-bed reactor system
was employed to treat dairy wastewater [93]. The maximum
COD removal rates obtained were between 93.8 and 98.5,
and 72.5 and 84%, respectively, for the two reactors
operated. The kinetic constants of anaerobic digestion of
dairy industry wastewater were determined using bioreac-
tors containing suspensions of micronized clays [94]. The
yield coefficient of methane (Yp) was computed to be
342 ml CH4/g COD. The system achieved around 90% COD
removal on average.
In another study, it was observed that bioconversion of
whey generated from cheese and casein manufacturing
process resulted in a pollutant reduction greater than 75%
[95]. The anaerobic treatment of salty cheese whey was
investigated using an anaerobic rotating biological contact
reactor [96]. The optimum performance was attained at an
HRT of 3 days and at 37 8C, resulting in a COD reduction of
85%. The methane content of the reactor biogas was around
74%. Application of an upflow anaerobic solid removal
(UASR) reactor was evaluated during pre-treatment of a
dairy wastewater [97]. At 20 8C and at 4.5 h of HRT, 98% of
lipids were removed at a pH of 4.0. A multichamber
anaerobic bioreactor was used to treat salty cheese whey
diluted with a mixed dairy wastewater [98]. The mesophilic
(37 8C) reactor provided a COD reduction of 83%, at an
HRT of 2 days. The methane content in the reactor biogas
was 68%. A non-attached bacterial growth process was
applied for biomethanation of a dairy wastewater [99]. In a
batch process, 92% COD reduction was obtained at 66 days.
The impacts of biofilm support systems on anaerobic
continuously stirred tank reactors (CSTRs) were described,
using a laboratory-scale reactor treating dairy wastewater
[100]. The authors reported that the incorporation of a
biofilm support system can significantly improve the
performance of the digester, due to development of active
biofilms that enhance biomass and waste contact, and reduce
the microbial wash-out. The process kinetics of the
mesophilic (37 8C) anaerobic digestion of synthetic ice-
cream wastewater were investigated at an HRT range
between 2.99 and 7.45 days, using the Monod and the
Contois equations, in a laboratory-scale study [101]. Since
the Contois equation incorporated the effect of any changes
in the influent substrate concentration, this kinetic model
described the process kinetic coefficients of the pilot-scale
anaerobic contact process better.
4. Two-phase anaerobic treatment of dairy wastewater
Two-phase anaerobic treatment systems are particularly
suitable for wastewaters containing high concentrations of
organic suspended solids, such as food and agricultural
industry wastewaters [102,103]. The performance of an acid
phase (acidogenic) reactor is especially of paramount
importance during two-phase anaerobic stabilization of
wastes, since the acid reactor should provide the most
appropriate substrate for the subsequent methane phase
(methanogenic) reactor. The guidelines for the design of pre-
acidification reactors for treating various high-strength
industrial wastewaters were already presented in literature
[104]. Actually, numerous studies have been performed
covering particularly the anaerobic acidogenesis of dairy
wastewaters. Initially, acidogenesis of lactose was investi-
gated extensively in laboratory-scale studies, mostly
focusing on the degradation kinetics of lactose [25–
28,105]. Studies covering the acid phase digestion of
industrial and synthetic dairy wastewaters have been
reviewed in different aspects previously in this paper
[10,11,14,15,22,24]. In one of these studies, anaerobic
digestion of three different dairy effluents (namely cheese,
fresh milk and milk powder/butter factories) was evaluated,
using a laboratory-scale mesophilic two-phase system [14].
For the cheese factory effluent, 97% COD removal was
achieved at an OLR of 2.82 kg COD/(m3 day), while at an
OLR of 2.44 kg COD/(m3 day), 94% COD removal was
obtained for the fresh milk effluent. For the powder milk/
butter factory effluent, 91% COD removal was achieved at
an OLR of 0.97 kg COD/(m3 day). In addition to these
works, the operating criteria for pre-acidification of dairy
wastewater obtained from a milk and cream bottling plant
were determined in a laboratory-scale CSTR [106]. The
maximum acidogenic conversion was computed to be 71%
by the authors. Acetic, propionic, n-butyric and n-valeric
acids were commonly produced during acidogenesis of dairy
wastewater. Anaerobic production of volatile fatty acids
(VFAs) with fermentation of whey permeate (wastewaters
from cheese-making process) was investigated in a
laboratory-scale anaerobic fluidized bed reactor [107]. It
has been shown that up to 19% of the initial sugar was
 B. Demirel et al. / Process Biochemistry 40 (2005) 2583–2595 2589

converted to volatile acids. Production of n-butyric acid was
also favoured during whey biodegradation. The influence of
substrate strength on thermophilic anaerobic acidogenesis of
a simulated dairy wastewater was investigated in a
laboratory-scale work [108]. In an influent substrate
concentration range between 2 and 30 g COD/l and at
55 8C, carbohydrate degraded under all conditions, however,
protein and lipid conversions both decreased when the
substrate concentration increased. The major acidogenesis
products were measured to be acetate, propionate, butyrate
and ethanol. Acidification of mid- and high-strength
synthetic dairy wastewaters were studied in a laboratory-
scale upflow reactor [109]. The authors reported that the
high-strength wastewater favoured production of hydrogen
(H2) and alcohols. Inhibitory effects of zinc (Zn) and copper
(Cu) on anaerobic acidogenesis of dairy wastewater were
investigated in a laboratory-scale study [110]. The authors
concluded that copper seemed more toxic than zinc to the
overall production of VFAs and hydrogen in the acidogenic
reactor. Start-up of two acidogenic reactors under meso-
philic (37 8C) and thermophilic (55 8C) conditions were
compared using a methanogenic granular sludge and dairy
wastewater [111]. It took more than 2 months to establish a
microbial community with a stable metabolic activity.
Acidogenesis of synthetic dairy wastewater was also studied
at a pH range between 4.0 and 6.5, in a laboratory-scale
upflow reactor at 37 8C [112]. At an HRT of 12 h and a pH of
5.5, 95% of carbohydrates, 82% of proteins and 41% of
lipids could be degraded in the acid phase reactor. Moreover,
batch reactors were used to investigate the thermophilic
anaerobic acidification of a synthetic dairy wastewater at a
pH of 5.5 [113]. According to the authors, hydrogen
production could be attributed to the fermentation of
carbohydrate. Two acidogenic upflow reactors were
operated under mesophilic (37 8C) and thermophilic
(55 8C) conditions with a synthetic dairy wastewater, in
order to compare the effects of temperature on the
performances of both reactors [114]. Almost no difference
was reported for the performances of both reactors in terms
of COD removal and degree of acidification, at any given
OLR. A cheese-processing wastewater was used to
determine the biokinetics of mesophilic acidogens [115].
At a pH of 7 and 36.2 8C, the maximum microbial growth
rate (mmax), half saturation coefficient (Ks), the microbial
yield coefficient (Y) and microbial decay rate (kd) were
computed to be 9.9 day 1, 134 mg COD/l, 0.29 mg MVSS/
mg COD and 0.14 day 1, respectively. While at a pH of 7.3
and 36.2 8C, mmax, Ks, Y and kd were determined to be
9.3 day 1, 482.5 mg COD/l, 0.20 mg MVSS/mg COD and
0.25 day 1, respectively. In a more recent study, the effects
of HRT between 12 and 24 h on anaerobic acidogenesis of
dairy wastewater was investigated, using a laboratory-scale
continuous-flow completely mixed anaerobic reactor with
solids recycle [116]. The acid production gradually
increased proportionally to the OLR, with decrease in
HRT. The highest degree of acidification and the rate of acid
production were 56% and 3.1 g/(l day), respectively, at 12 h
of HRT.
A summary of data for two-phase anaerobic dairy waste-
water treatment practices in literature is given in Table 4.
Initial results for two-phase anaerobic treatment of a dairy
wastewater were reported from pilot plant studies [117]. The
authors reported that the fluidized bed reactor configuration
was best suited to the two-phase operation, since this process
could achieve a high concentration of biomass in the reactor
without the need for the bacterial separation between the
stages. Two-phase anaerobic treatment of dilute milk wastes
was investigated, using a CSTR for acidogenesis and an
upflow filter reactor for methanogenesis, respectively [118].
The system attained 92% COD removal at an overall HRT of
4.4 days for an influent substrate concentration of
1500 mg COD/l. Anaerobic fluidized beds were also used
in another work, in order to treat a dairy effluent with an
influent COD concentration of 5000 mg COD/l [119].

Table 4
Two-phase anaerobic treatment process performances for dairy wastewater treatment
Waste type Reactor type HRT Loading Temperature Methane yield Removal Application Reference
(8C) (m3/kg CODr) (%) status
Cheese-fresh milk-powder 2.82–2.44–0.97 35 0.359–0.327 97–94–91 Laboratory [14]
milk/butter effluents (kg COD/(m3 day)) –0.287 (COD) scale
Dilute milk waste CSTR + upflow 4.4 (day) 92 (COD) [117]
filter
Fluidized beds 9.4 (kg COD/ 76–92 (COD) [118]
(m3 day))
Wastewater of a milk CSTR + upflow 2 (day) 35 90 (COD) Laboratory [120]
bottling plant filter scale
Skimmed milk CSTR + upflow 2 (day) 20 95.5 (COD) Laboratory [121]
filter scale
Wastewater of a milk and CSTR + upflow 2 (day) 5 (kg COD/ 33–36 90 (COD), Laboratory [122]
cream bottling plant filter (m3 day)) 95 (BOD) scale
Synthetic cheese whey CSTR + upflow 5 (day) 35 95 (COD) Laboratory [123]
filter scale
2590 B. Demirel et al. / Process Biochemistry 40 (2005) 2583–2595

Substrate removal efficiencies varied between 76 and 92%, at
an OLR of 9.4 kg COD/(m3 day) in the two-phase system.
Anaerobic treatment of cheese whey was investigated in a
pilot-scale study [120]. The UASB reactors were used as
acidogenic and methanogenic reactors in this work. Two-
phase anaerobic digestion process for dairy wastewater
treatment was also examined from a microbiological point of
view during start-up in a laboratory-scale study [121]. The
authors reported that the numbers of acidogens remained
constant in the pre-acidification reactor (a CSTR), while the
numbers of methanogens and non-methanogens slightly
decreased in the methanogenic upflow anaerobic filter reactor.
Treatment characteristics of two different substrates, baby
nutrient milk and skimmed milk, were investigated using a
two-phase anaerobic digestion process at 20 8C [122]. At an
HRT of 2 days, 96% COD reduction was obtained for
skimmed milk with an influent substrate concentration of
1500 mg COD/l. The performance of a laboratory-scale
mesophilic two-phase anaerobic digestion system treating
dairy wastewater from a milk bottling plant was evaluated,
using a CSTR for acidogenesis and an upflow anaerobic filter
for methanogenesis [123]. The system achieved overall COD
and BOD removal rates of 90 and 95%, respectively, at an
HRT of 2 days and an OLR of 5 kg COD/(m3 day). In another
laboratory-scale study, a CSTR for acidogenesis and an
upflow anaerobic filter for methanogenesis were used at
35 8C, for two-phase anaerobic treatment of cheese whey
[124]. In the upflow methanogenic filter, 95% COD removal
was attained at an HRT of 4 days, with a biogas production of
0.55 m3/kg COD removed. Changes to bacterial community
in a laboratory-scale two-phase anaerobic digestion system
treating dairy wastewater was reported in a recent study [125].
The ratio of total autofluorescent methanogens to total
bacteria varied between 0.01 and 3% in the acid phase reactor
(a CSTR), and 2–13% in the methane phase reactor (an upflow
anaerobic filter). The system provided a methane yield
between 0.30 and 0.34 m3 CH4/kg COD removed during the
entire operation.
In both conventional—single and two-phase anaerobic
treatment processes, primary objectives are consistent:
achievement of a high degree of waste stabilization and a
high conversion of waste to methane. Furthermore, for a
particular type of wastewater, in order to achieve these goals
specified, the maximum loadings applicable and possible
treatment rates for a particular reactor configuration are
obviously also important. For anaerobic treatment of dairy
wastewaters, the maximum loadings and corresponding
treatment rates achieved with various reactor configurations
in literature are briefly summarized in Table 5. Typical
operating conditions for anaerobic digesters are also
outlined in Table 6 [2]. High influent substrate concentra-
tions, fluctuations in dairy wastewater flow rate and

Table 5
Maximum loadings and treatment rates in anaerobic treatment of dairy wastewater
Process mode Application status Wastewater type Reactor type Maximum loading Maximum removal Reference
3
Single phase Pilot plant Milk and cream Upflow filter 6 kg COD/(m day) >85% COD [4]
Two phase Laboratory scale Cheese Hybrid 2.82 kg COD/(m3 day) 97% COD [14]
Single phase Laboratory scale Cheese UASB 7.5 g COD/(l day) 85–90% COD [17]
Single phase Whey Semicontinuous 16.1 kg COD/(m3 day) >99% COD [54]
digester
Single phase Upflow filter 21 kg COD/(m3 day) [65]
Single phase Pilot plant Ice-cream Upflow filter 18 kg COD/(m3 day) [66]
Single phase Laboratory scale Cheese whey Upflow filter >20 kg COD/(m3 day) [71]
Single phase Laboratory scale Raw milk quality Upflow filter 5–6 kg COD/(m3 day) >90% COD [73]
control laboratory
Single phase Laboratory scale Cheese whey UASB 38.1 g COD/l [77]
(as influent concentration)
Single phase Laboratory scale Cheese UASB >45 g COD/(l day) 70–80% COD [78]
Single phase Laboratory scale UASB 10.8 kg COD/(m3 day) [85]
Single phase Laboratory scale Cheese whey Hybrid bed 11 kg COD/(m3 day) >95% COD [87]
Single phase Anaerobic SBR 6 g COD/(l day) [91]
Two phase Fluidized bed 9.4 kg COD/(m3 day) 76–92% COD [119]
Two phase Laboratory scale Milk and cream CSTR + upflow filter 23–7 kg COD/(m3 day) 90% COD, 95% BOD5 [123]
bottling

Table 6
Typical operating conditions for anaerobic digesters [2]
Anaerobic digester configuration Load (kg COD/(m3 day)) Retention COD removal (%)
CSTR 0.5–2.5 1–5 days 80–90
Anaerobic filter 2–10 10–50 h 70–80
UASB 2–15 8–50 h 70–90
Fluidized bed 2–50 0.5–24 h 70–80
 B. Demirel et al. / Process Biochemistry 40 (2005) 2583–2595
composition, excessive suspended solids and lipid concen-
trations in dairy effluents, the presence of sufficient amount
of alkalinity in anaerobic digesters and appropriate
anaerobic reactor configuration employed for treatment
are common factors that affect maximum loading rates and
expected treatment efficiencies [4,14,66,73,77,78,81,116,
123]. High variability in composition and flow rate of dairy
wastewaters are probably the most important parameters that
should be considered initially. Thus, installing an equaliza-
tion basin prior to anaerobic/aerobic treatment facility is
usually required for stable process efficiency. High
suspended solids concentrations in dairy effluents particu-
larly affect treatment performances of anaerobic CSTRs and
filters [2,4]. High substrate concentrations were reported to
decrease UASB reactor performance treating dairy waste-
water [77,81]. Biomass granulation must definitely be
achieved for stable and satisfactory UASB reactor operation
[76,78]. Moreover, dairy wastewaters usually contain
differing proportions of fats and proteins that require longer
time for hydrolysis [101]. Therefore, larger reactor volumes
may be required to treat larger volumes of wastewater at
high HRTs for such dairy effluents. Presence of lipids in
single-phase anaerobic filter treatment for dairy effluents is
also a common problem, because anaerobic filters remove
lipids by entrapment, without biodegradation. This may
soon result in channelling and clogging, with a subsequent
decrease in reactor performance. Choice of appropriate
packing materials in upflow anaerobic filters also affect
maximum substrate loading rates and expected treatment
rates significantly [63,65]. Poor biodegradation of milk-fat
was attributed to the limiting rate of liquefaction, indicated
by a low hydrolysis constant in an expanded granular sludge
bed (EGSB) reactor [36]. Utilization of two-phase anaerobic
treatment processes seems a favourable choice to overcome
suspended solids and particularly lipid problems for
concentrated dairy wastewaters [122]. Besides, a two-phase
system will protect the methanogens in the methane reactor
from inhibitory levels of pH and high amounts of VFAs
produced in the first-acid reactor. Acidogenic phase will
Table 7
Anaerobic/aerobic treatment performance levels for dairy wastewaters
Effluent type System configuration
Milk bottling plant DAF + upflow anaerobic filter (UAF)
Cheese whey Downflow–upflow hybrid
anaerobic reactor (DUHR) + SBR
Cheese wastewater UASB pond + aerated pond
Synthetic milk powder/butter AAO activated sludge
factory wastewater
Wastewater from an industrial Anaerobic filter + SBR
milk analysis laboratory
2591
allow pH control in the first phase, if required, in a pilot or
full-scale treatment plant [86]. Finally, sufficient amount of
alkalinity is frequently discussed to be the most important
factor for controlling process reliability during anaerobic
treatment of dairy wastewater [66,73,78,87].
5. Anaerobic/aerobic treatment of dairy wastewater
Aerobic treatment processes are commonly used together
with anaerobic processes for dairy wastewater treatment, in
order to achieve the effluent discharge limits for agro-industry
wastewaters. Results for anaerobic/aerobic treatment of dairy
wastewaters are outlined in Table 7. Among the studies
mentioned in this paper, pre-treatment of a dairy wastewater
from a milk bottling plant was reported using a pilot-scale
dissolved air floatation (DAF) unit and a pilot-scale anaerobic
upflow filter reactor [4]. The aim was to attain a BOD5
reduction of between 38 and 50% and a SS reduction of 60–
75% in the DAF unit prior to the biological treatment step. The
upflow anaerobic filter reactor achieved BOD5 and COD
reduction rates greater than 90 and 85%, respectively, with a
biogas yield of 0.40 l biogas/g COD removed. A laboratory-
scale anaerobic–aerobic biological process was used to treat
cheese whey [12]. The anaerobic downflow–upflow hybrid
reactor (DUHR) achieved 98% COD reduction at an OLR of
10 g COD/(l day). Post treatment was subsequently per-
formed using a SBR, resulting in more than 90% of both COD
and nutrient removal rates. Furthermore, full-scale anaerobic/
aerobic treatment of cheese wastewater by a system
comprising a grease trap, an UASB type pond, an aerated
pond and an effluent polishing pond was reported [13].
Reduction rates in BOD5, COD, TSS, and oil and grease were
98, 96, 98 and 99.8%, respectively. In addition to these
findings, a low cost treatment system was proposed to reduce
the strength of a dairy waste [126]. This three-stage treatment
system consisted of a sump in which the anaerobic digestion
takes place, an aerobic vegetated filter, and an irrigated
plantation. Biological phosphorus removal from a cheese
Removal Application status Reference
38–50% BOD5 (DAF)
>90% BOD5 (UAF) Pilot scale [4]
>85% COD (UAF)
98% COD (DUHR)
>90% COD (SBR) Laboratory scale [12]
98% (BOD5)
96% (COD) Full scale [13]
98% (TSS)
>90% (COD) Laboratory scale [127]
98% (COD), 99% (nitrogen) [131]
2592 B. Demirel et al. / Process Biochemistry 40 (2005) 2583–2595
factory effluent was reported using a bench-scale SBR [127].
The existing industrial-scale treatment facility at the factory
consisted of an anaerobic equalization tank, followed by an
UASB and aerated lagoons. The effluent orthophosphates
concentration was around 5 mg/l in the bench-scale SBR
effluent, pointing out a better treatment result than that of the
industrial-scale treatment facility. A laboratory-scale acti-
vated sludge system was used to treat synthetic wastewater
from a milk powder/butter factory [128]. The substrate is fed
to anaerobic and anoxic sectors, in series with an aerobic
reactor, while the sludge is returned to the anaerobic selector,
and the mixed liquor from the aerobic sector is recycled to the
anoxic sector. Over 90% COD removal efficiency could be
achieved at an HRTof 7 days and at a nominal sludge age of 20
days in the system. Treatment of a high-strength dairy
wastewater was reported using a low-rate anaerobic pre-
treatment process and an aerobic polishing step, in an HRT
range of 5–7 days in a pilot-scale study [129]. Microbial
removal from dairy wastewater was also reported recently,
using a combined treatment system consisting of paired solids
separators, anaerobic lagoons, aerobic ponds and constructed
wetlands cells [130]. The authors reported that high turbidity
levels in dairy waste stream decreased the capability of the
treatment system, in terms of removal of some microbial
indicators and pathogens. Wastewaters from an industrial
milk analysis laboratory were treated using an anaerobic filter
(AF) and a SBR operated in series [131]. Effluent soluble
COD and total nitrogen concentrations were below 200 and
10 mg/l, respectively. Moreover, the authors concluded that
the combination of an AF and a SBR resulted in a lower
energy consumption and sludge generation. A wheat straw
biofilter was operated in a sequential aerobic–anaerobic mode
in a temperature range between 8 and 14 8C, to treat
wastewater from a milkhouse and milking parlour [132]. The
attenuation of pollutants in dairy wastewater for TSS, oil and
grease, and COD were determined to be 89, 76 and 37%,
respectively. Biological phosphorus removal from a synthetic
phosphorus-rich dairy wastewater was evaluated using an
anaerobic reactor and an activated sludge reactor [133]. The
system resulted in a final sludge phosphorus content of 4.9%
mg P/mg TSS. Utilization of aerobic/anaerobic membrane
bioreactors coupled with anaerobic digestion seemed to be a
feasible method for treating wastewaters from livestock
operations, such as dairy wastewater [134]. It was reported
that high-quality reusable water can be produced using
membrane bioreactors in the treatment of such wastewaters.
Recently, biological treatment of dairy wastewater was
evaluated in a laboratory-scale work, using anaerobic and
aerobic sequencing batch reactors [135]. The SBR system was
found to result in effective denitrification by the authors.
6. Conclusions
Conventional anaerobic treatment processes are often used
for treating dairy wastewaters. Particularly anaerobic filters
and UASB reactors are the most common reactor configura-
tions employed. In fact, the UASB reactors are very suitable
for treating food industry wastewaters, since they can treat
large volumes of wastewaters in a relatively short period of
time. More research should be directed towards treatment of
dairy wastewaters in pilot and full-scale UASB reactors in
near future, to make use of these potential advantages
outlined. Lipid degradation and inhibition in single-phase
anaerobic systems is frequently discussed in literature, since
lipids are potential inhibitors in anaerobic systems, which can
often be encountered by environmental engineers and
wastewater treatment plant operators. Moreover, high
concentrations of suspended solids in dairy waste streams
can also affect the performance of conventional anaerobic
treatment processes adversely, particularly the most com-
monly used upflow anaerobic filters. Thus, two-phase
anaerobic digestion processes should be considered more
often to overcome these problems that may be experienced in
conventional single-phase design applications, since two-
phase anaerobic treatment systems are reported to produce
better results with various industrial wastewaters, such as
olive oil mill and food-processing effluents, which are high in
suspended solids and lipids content. When two-phase
anaerobic digestion processes are evaluated as a whole, it
is clear that the acid phase digestion of dairy wastewaters is
actually investigated in various aspects. However, data
especially for full-scale two-phase applications for dairy
effluents in literature is scarce. Furthermore, in addition to
degradation of lipids, protein solubilization should be
investigated more comprehensively during acid phase
digestion of dairy wastes with a relatively high protein
content, because there is contradictory information in
literature about protein solubilization with different waste-
water types during anaerobic acidogenesis. Since high rate
anaerobic treatment of dairy wastes (or any industrial
wastewater) with a relatively higher content of particulates,
fats and proteins can often be problematic, modelling studies
simulating biodegradation mechanisms of these components
should extensively be explored. Removal of nitrogen and
phosphorus from dairy wastewaters has recently gained
significant attention, due to more strict environmental
regulations, so current research efforts clearly seem to focus
on this particular topic. Recently, bench–pilot and full-scale
applications of combined treatment methods for nutrient
removal from dairy waste effluents are frequently encoun-
tered. It is obvious that as the regulations for discharge of
nutrients become more strict in time, new modifications in
existing treatment plants will eventually be incorporated.
Finally, since the anaerobic digestion process is an imperative
tool for the production of clean energy sources, such as
hydrogen and methane, biogas production from high-strength
dairy industry wastes will always be of paramount
importance, as a valuable renewable energy source, for both
developed and developing countries in future. Particularly,
production of hydrogen by acidogenesis of high-strength
dairy waste effluents is currently worth investigating.
 B. Demirel et al. / Process Biochemistry 40 (2005) 2583–2595
Acknowledgements
The authors wish to express their gratitude to the financial
support by the Bogazici University Research Fund through
project number 01Y101D.
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