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Process Biochemistry 40 (2005) 25832595 www.elsevier.

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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 efuents. 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 identied 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 benchpilotfull-scale anaerobic treatment efforts for dairy waste efuents. Combined (anaerobicaerobic) treatment methods are also discussed. Finally, areas where further research and attention are required are identied. # 2005 Elsevier Ltd. All rights reserved.
Keywords: Anaerobic treatment; Dairy wastewaters; Acidogenesis; Lipids degradation; Proteins degradation

1. Introduction The dairy industry, like most other agro-industries, generates strong wastewaters characterized by high biological 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 efuents in Europe. A typical European dairy generates approximately 500 m3 of waste efuent daily [2]. Dairy waste efuents are concentrated in nature, and the main contributors of organic load to these efuents are carbohydrates, proteins and fats originating from the milk [3,4]. Since dairy waste streams contain high concentrations of organic matter, these efuents 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). 0032-9592/$ see front matter # 2005 Elsevier Ltd. All rights reserved. doi:10.1016/j.procbio.2004.12.015

malfunctions or operational errors [4,6]. Dairy wastewaters are treated using physico-chemical and biological treatment methods. However, since the reagent costs are high and the soluble COD removal is poor in physicalchemical 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 signicant drawback of these processes. COD concentrations of dairy efuents vary signicantly; moreover, dairy efuents 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 efuents. In the paper, the general characteristics of dairy waste streams are identied 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 (anaerobicaerobic)

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treatment systems for dairy wastewaters are also summarized briey. Finally, areas where particular research and more attention required in the near future are identied.

2. General characteristics of dairy wastewaters Wastewaters from the dairy industry are usually generated in an intermittent way, so the ow rates of these efuents change signicantly. 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 efuents 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 inuences 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 operations in literature is scarce. Only one comprehensive study has been encountered, which provides extensive information
Table 1 Characteristics of dairy waste efuents Efuent type Creamery Not given Mixed dairy processing Cheese whey Cheese Fresh milk Cheese Milk powder/ butter Mixed dairy processing Cheese whey Cheese Not given Fluid milk
a

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 efuents from full-scale operations is given in Table 1 [11 19]. High COD concentrations indicate that dairy industry wastewaters are strong and uctuating in nature. Signicant 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 polyphosphate (P2O74), as well as organic forms [20]. Concentrations 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 nes or avoring 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

COD (mg/l) 20006000 9807500 11509200 68814a 10007500 4656a 5340a 1908a 63100a 61000a

BOD5 (mg/l) 12004000 6804500

pH (units) 811 611

Alkalinity (mg CaCO3/l) 150300 320970

Suspended solids (mg/l) 3501000 300 3401730

Volatile suspended solids (mg/l) 330940 255830

Total solids (mg/l)

TKN (mg/l) 5060

Total phosphorus (mg/l)

Reference

27053715

14272 1462a

868 379a

[4] [9] [11] [12] [13] [14] [14] [14] [15]

5885000

5.59.5 6.92a 5.22a 5.80a 3.35a

5002500

12500a 1780a 2500a 90450 90450

12100a 1560a

53000a 980a 830a 510a 280a

9502400

5001300

4.7a 4.49.4 5.09.5

[16] [17] [18] [19]

Mean concentrations are reported.

Table 2 Concentrations of selected elements in dairy wastewaters Efuent type Creamery Cheese/whey Cheese/alcohol Cheese/beverages Cheese/whey Mixed dairy Cheese
a

Na (mg/l) 170200 735a 423a 453a 419a 1232324 720980

K (mg/l) 3540 42.8a 41.2a 8.6a 35.8a 8160

Ca (mg/l) 3540 47.7a 54.3a 33.6a 52.3a 12120 530950

Mg (mg/l) 58 11.4a 8.3a 16.9a 11.0a 297

Fe (mg/l) 25

Co (mg/l) 0.050.15

Ni (mg/l) 0.51.0

Mn (mg/l) 0.020.10

Reference [4] [6] [6] [6] [6] [11] [13]

0.56.7

00.13

0.030.43

Mean concentrations are reported.

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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 dened as a complex type of substrate [2224]. Lactose is the main carbohydrate in dairy wastewater and is a readily available substrate for anaerobic bacteria. Anaerobic methanation 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 ow schemes were proposed for acidogenic fermentation of lactose; carbon ow 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 efuents. 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 rstly hydrolyzed to glycerol and long chain fatty acids (LCFAs), followed by b-oxidation, producing acetate and hydrogen [29]. The biodegradation of lipids is difcult 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 conventional 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]. Difculties experienced with the presence of lipids in anaerobic treatment processes have been previously reported in literature[4144].

3. Conventional (single-phase) anaerobic treatment of dairy efuents Anaerobic treatment processes are favourable methods for treating dairy waste efuents, in comparison to aerobic processes, due to their well-known benets for treating industrial wastewaters, particularly from agricultural industries with a high organic content [4547]. Anaerobic treatment applications for dairy industry wastewaters have been evaluated in a number of previous studies [4859]. More recent information about anaerobic treatment practices of dairy waste streams are also displayed in Table 3. In treatment studies of dairy wastewaters, anaerobic lters have recently been used. If the particular dairy efuents contain low concentrations of suspended solids, then anaerobic lter reactors are generally suitable for biological treatment. A laboratory-scale plastic medium anaerobic lter 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 lters treating dairy wastewater, in terms of COD removal, was not signicantly affected by temperature variations between 21 and 30 8C [62]. The effects of porous and non-porous support media in anaerobic lter reactors on thermophilic anaerobic treatment of ice-cream wastes were investigated extensively [63,64]. At high loading rates, anaerobic lter with porous support media performed more satisfactorily [63]. The performance of porous and non-porous media in an upow anaerobic lter (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 lter 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 lters 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 rst-order rate constant for each anaerobic lter reactor. In another work, an upow anaerobic lter 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, efuent SS and COD concentrations satised the efuent discharge limits. A pilotscale upow anaerobic lter reactor treating dairy wastewaters 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

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Table 3 Anaerobic treatment performance levels for dairy wastewaters Waste type Cheese whey Cheese whey Ice-cream Reactor type Downow xed-lm Downow xed-lm Anaerobic lter Anaerobic lter Anaerobic lter Anaerobic lter Anaerobic lter UASB UASB UASB Hybrid UASB Hybrid UASB Cheese production wastewater UASB UASB Synthetic dairy efuent Cheese whey Synthetic non-fat dry milk Hybrid Hybrid ASBR ASBR Upow packed-bed Washing and rinsing waters Cheese whey Rotating biological contact reactor Upow anaerobic solid removal reactor Multichamber bioreactor CSTR 3 (day) 4.5 (h) 2 (day) 7.455.994.60 3.762.99 (day) 37 20 37 35 6 1020 3040 (day) 4.11.7 (day) 2 (day) 6 (h) 36 (day) 24 (g VS/(l day)) 514.29 (kg COD/(m3 day)) 0.45 (day) 5 (h) 8.5 (g COD/(l day)) 18 (g COD/(l day)) 27.3 2.34.5 1.51.9 (g COD/(l day)) 0.826.11 (kg COD/(m day)) Up to 11 (kg COD/(m3 day)) 5
3

HRT 4.9 (day) 6.6 (day) 4 (day)

Loading 13 (kg COD/(m3 day)) 8.3 (kg COD/(m3 day))

Temperature (8C)

Methane yield (m3/kg CODra) 0.28 0.34

Removal (%) 75 (COD) 76 (COD) 928578 (COD) 75 (COD) 85 (COD) 80 (COD) 90 (COD) >97 (COD)

Application status Pilot scale Pilot scale Laboratory scale Pilot scale Pilot scale Full scale Laboratory scale Laboratory scale Laboratory scale Laboratory scale Laboratory scale

Reference [16] [16] [60,61] [66] [69] [72] [73] [75] B. Demirel et al. / Process Biochemistry 40 (2005) 25832595 [78] [80] [19] [82]

302112.5 5.5 (kg COD/(m3 day)) 6 (kg COD/(m3 day)) Up to 21 (kg COD/(m3 day)) 56 (kg COD/(m3 day)) 0.320.34

0.5 (day)

Raw milk Cheese whey Cheese production wastewater Whey permeate Efuent of an integrated plant

31 (g COD/(l day))

90 (COD) 9964.2 (COD) 30 30 87 (COD) 92 8599 35 7991 81 90 (COD) 0.354 (at 1.7 days HRT) 9097 (COD) >95 (COD) 62 (COD), 75 (BOD5) 2650 (VS) 93.898.5, 72.584 (COD) 90.4 (COD) 85 (COD) 98 (lipid) 83 (COD) 9897969492 (SCOD)b

Laboratory scale

[17] [83]

35

Laboratory scale Laboratory scale Laboratory scale Laboratory scale Laboratory scale

[85] [86] [87] [88,89] [92] [93] [95]

Laboratory scale

[96] [98]

Cheese whey Synthetic ice-cream wastewater


a b

Laboratory scale

[100]

COD removal. Soluble COD.

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non-staged anaerobic lters treating synthetic dairy wastewaters 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. Laboratoryscale single-fed (SFR) and multi-fed (MFR) upow anaerobic lters 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 specic activities of the different trophic groups were found to be higher in the MFR. An upow anaerobic lter 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 lter 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 lter reactor. Upow 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 inuent 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 efuents 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 specic 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 inuent 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 coefcients of laboratory-scale continuous UASB reactors treating whey permeate were determined, in an HRT range between 0.4 and 5 days and at a

constant inuent substrate concentration of 10.4 0.2 g COD/l [79]. The maximum substrate utilization rate (k), half saturation coefcient (KL), yield coefcient (Y) and decay rate coefcient (Kd) were determined to be 0.941, 0.773 kg COD/(kg VSS day), 0.153 kg VSS/(kg COD) and 0.022 day1, respectively. Under the same HRT and inuent substrate concentration, COD removal efciency 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 inuent in an UASB reactor [81]. For proper system stability, the optimum inuent 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 ow mixing chamber of an anaerobic lter 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 upow liquid and gas velocities, which enables treatment of low strength wastes at short HRTs, as well as treatment of high-strength efuents 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 9290%, for both reactors. In addition to anaerobic lters and UASB reactors, hybrid digesters and anaerobic sequencing batch reactors (ASBR) are also employed for treating dairy efuents. A mesophilic laboratory-scale hybrid anaerobic digester, combining an upow sludge blanket and a xed-bed design, was used to treat synthetic dairy efuent with an inuent 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 upow hybrid reactor resulted in removal efciencies 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 efciencies for dairy efuents. 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

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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 2644%, 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 24 g VS/(l day) and at HRTs of 3 and 6 days. The purication 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 purication efciency 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 lter, anaerobic expanded/uidized bed reactor and UASB reactor, all treating ice-cream wastewater, were comprehensively examined during start-up [92]. The authors reported that the reactor conguration did not play an important role in causing changes in the microbial community. A two-stage upow 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 bioreactors containing suspensions of micronized clays [94]. The yield coefcient 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 upow 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 biolm 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

biolm support system can signicantly improve the performance of the digester, due to development of active biolms that enhance biomass and waste contact, and reduce the microbial wash-out. The process kinetics of the mesophilic (37 8C) anaerobic digestion of synthetic icecream 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 inuent substrate concentration, this kinetic model described the process kinetic coefcients 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 preacidication 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 investigated 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 efuents (namely cheese, fresh milk and milk powder/butter factories) was evaluated, using a laboratory-scale mesophilic two-phase system [14]. For the cheese factory efuent, 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 efuent. For the powder milk/ butter factory efuent, 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-acidication 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 uidized bed reactor [107]. It has been shown that up to 19% of the initial sugar was

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converted to volatile acids. Production of n-butyric acid was also favoured during whey biodegradation. The inuence of substrate strength on thermophilic anaerobic acidogenesis of a simulated dairy wastewater was investigated in a laboratory-scale work [108]. In an inuent 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. Acidication of mid- and high-strength synthetic dairy wastewaters were studied in a laboratoryscale upow 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 mesophilic (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 upow 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 acidication 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 upow 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 acidication, 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 coefcient (Ks), the microbial yield coefcient (Y) and microbial decay rate (kd) were computed to be 9.9 day1, 134 mg COD/l, 0.29 mg MVSS/ mg COD and 0.14 day1, respectively. While at a pH of 7.3 and 36.2 8C, mmax, Ks, Y and kd were determined to be 9.3 day1, 482.5 mg COD/l, 0.20 mg MVSS/mg COD and 0.25 day1, 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-ow 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 acidication 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 wastewater 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 uidized bed reactor conguration 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 upow lter reactor for methanogenesis, respectively [118]. The system attained 92% COD removal at an overall HRT of 4.4 days for an inuent substrate concentration of 1500 mg COD/l. Anaerobic uidized beds were also used in another work, in order to treat a dairy efuent with an inuent COD concentration of 5000 mg COD/l [119].

Table 4 Two-phase anaerobic treatment process performances for dairy wastewater treatment Waste type Cheese-fresh milk-powder milk/butter efuents Dilute milk waste CSTR + upow lter Fluidized beds CSTR + upow lter CSTR + upow lter CSTR + upow lter CSTR + upow lter 4.4 (day) 9.4 (kg COD/ (m3 day)) 2 (day) 2 (day) 2 (day) 5 (day) 5 (kg COD/ (m3 day)) 35 20 3336 35 Reactor type HRT Loading 2.822.440.97 (kg COD/(m3 day)) Temperature (8C) 35 Methane yield (m3/kg CODr) 0.3590.327 0.287 Removal (%) 979491 (COD) 92 (COD) 7692 (COD) 90 (COD) 95.5 (COD) 90 (COD), 95 (BOD) 95 (COD) Laboratory scale Laboratory scale Laboratory scale Laboratory scale Application status Laboratory scale Reference [14] [117] [118] [120] [121] [122] [123]

Wastewater of a milk bottling plant Skimmed milk Wastewater of a milk and cream bottling plant Synthetic cheese whey

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Substrate removal efciencies 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. Twophase 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-acidication reactor (a CSTR), while the numbers of methanogens and non-methanogens slightly decreased in the methanogenic upow anaerobic lter 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 inuent 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 upow anaerobic lter 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 upow anaerobic lter for methanogenesis were used at

35 8C, for two-phase anaerobic treatment of cheese whey [124]. In the upow methanogenic lter, 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 autouorescent methanogens to total bacteria varied between 0.01 and 3% in the acid phase reactor (a CSTR), and 213% in the methane phase reactor (an upow anaerobic lter). The system provided a methane yield between 0.30 and 0.34 m3 CH4/kg COD removed during the entire operation. In both conventionalsingle 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 specied, the maximum loadings applicable and possible treatment rates for a particular reactor conguration are obviously also important. For anaerobic treatment of dairy wastewaters, the maximum loadings and corresponding treatment rates achieved with various reactor congurations in literature are briey summarized in Table 5. Typical operating conditions for anaerobic digesters are also outlined in Table 6 [2]. High inuent substrate concentrations, uctuations in dairy wastewater ow rate and

Table 5 Maximum loadings and treatment rates in anaerobic treatment of dairy wastewater Process mode Single phase Two phase Single phase Single phase Single Single Single Single phase phase phase phase Application status Pilot plant Laboratory scale Laboratory scale Wastewater type Milk and cream Cheese Cheese Whey Reactor type Upow lter Hybrid UASB Semicontinuous digester Upow lter Upow lter Upow lter Upow lter UASB UASB UASB Hybrid bed Anaerobic SBR Fluidized bed CSTR + upow lter Maximum loading 6 kg COD/(m day) 2.82 kg COD/(m3 day) 7.5 g COD/(l day) 16.1 kg COD/(m3 day) 21 kg COD/(m3 day) 18 kg COD/(m3 day) >20 kg COD/(m3 day) 56 kg COD/(m3 day) 38.1 g COD/l (as inuent concentration) >45 g COD/(l day) 10.8 kg COD/(m3 day) 11 kg COD/(m3 day) 6 g COD/(l day) 9.4 kg COD/(m3 day) 237 kg COD/(m3 day)
3

Maximum removal >85% COD 97% COD 8590% COD >99% COD

Reference [4] [14] [17] [54] [65] [66] [71] [73] [77]

Pilot plant Laboratory scale Laboratory scale Laboratory scale Laboratory scale Laboratory scale Laboratory scale

Single phase Single phase Single phase Single phase Single phase Two phase Two phase

Ice-cream Cheese whey Raw milk quality control laboratory Cheese whey Cheese Cheese whey

>90% COD

7080% COD >95% COD 7692% COD 90% COD, 95% BOD5

Laboratory scale

Milk and cream bottling

[78] [85] [87] [91] [119] [123]

Table 6 Typical operating conditions for anaerobic digesters [2] Anaerobic digester conguration CSTR Anaerobic lter UASB Fluidized bed Load (kg COD/(m3 day)) 0.52.5 210 215 250 Retention 15 days 1050 h 850 h 0.524 h COD removal (%) 8090 7080 7090 7080

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composition, excessive suspended solids and lipid concentrations in dairy efuents, the presence of sufcient amount of alkalinity in anaerobic digesters and appropriate anaerobic reactor conguration employed for treatment are common factors that affect maximum loading rates and expected treatment efciencies [4,14,66,73,77,78,81,116, 123]. High variability in composition and ow rate of dairy wastewaters are probably the most important parameters that should be considered initially. Thus, installing an equalization basin prior to anaerobic/aerobic treatment facility is usually required for stable process efciency. High suspended solids concentrations in dairy efuents particularly affect treatment performances of anaerobic CSTRs and lters [2,4]. High substrate concentrations were reported to decrease UASB reactor performance treating dairy wastewater [77,81]. Biomass granulation must denitely 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 efuents. Presence of lipids in single-phase anaerobic lter treatment for dairy efuents is also a common problem, because anaerobic lters 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 upow anaerobic lters also affect maximum substrate loading rates and expected treatment rates signicantly [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 rst-acid reactor. Acidogenic phase will
Table 7 Anaerobic/aerobic treatment performance levels for dairy wastewaters Efuent type Milk bottling plant System conguration DAF + upow anaerobic lter (UAF)

allow pH control in the rst phase, if required, in a pilot or full-scale treatment plant [86]. Finally, sufcient 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 efuent 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 oatation (DAF) unit and a pilot-scale anaerobic upow lter 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 upow anaerobic lter 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 laboratoryscale anaerobicaerobic biological process was used to treat cheese whey [12]. The anaerobic downowupow hybrid reactor (DUHR) achieved 98% COD reduction at an OLR of 10 g COD/(l day). Post treatment was subsequently performed 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 efuent 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 ndings, 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 lter, and an irrigated plantation. Biological phosphorus removal from a cheese

Removal 3850% BOD5 (DAF) >90% BOD5 (UAF) >85% COD (UAF) 98% COD (DUHR) >90% COD (SBR)

Application status Pilot scale

Reference [4]

Cheese whey

Downowupow hybrid anaerobic reactor (DUHR) + SBR UASB pond + aerated pond

Laboratory scale Full scale Laboratory scale

[12] [13] [127] [131]

Cheese wastewater

98% (BOD5) 96% (COD) 98% (TSS) >90% (COD) 98% (COD), 99% (nitrogen)

Synthetic milk powder/butter factory wastewater Wastewater from an industrial milk analysis laboratory

AAO activated sludge Anaerobic lter + SBR

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factory efuent 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 efuent orthophosphates concentration was around 5 mg/l in the bench-scale SBR efuent, pointing out a better treatment result than that of the industrial-scale treatment facility. A laboratory-scale activated 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 efciency 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 pretreatment process and an aerobic polishing step, in an HRT range of 57 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 lter (AF) and a SBR operated in series [131]. Efuent 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 biolter was operated in a sequential aerobicanaerobic 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 nal 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 denitrication by the authors.

6. Conclusions Conventional anaerobic treatment processes are often used for treating dairy wastewaters. Particularly anaerobic lters

and UASB reactors are the most common reactor congurations 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 commonly used upow anaerobic lters. 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 twophase anaerobic treatment systems are reported to produce better results with various industrial wastewaters, such as olive oil mill and food-processing efuents, 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 efuents 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 wastewater 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 signicant attention, due to more strict environmental regulations, so current research efforts clearly seem to focus on this particular topic. Recently, benchpilot and full-scale applications of combined treatment methods for nutrient removal from dairy waste efuents are frequently encountered. It is obvious that as the regulations for discharge of nutrients become more strict in time, new modications 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 efuents is currently worth investigating.

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Acknowledgements The authors wish to express their gratitude to the nancial support by the Bogazici University Research Fund through project number 01Y101D.

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