Dairy Industry Effluents Treatment

Anaerobic Treatment of Whey in Stirred Batch Reactor Abdulrzzak Alturkmani, Dr. Eng., Technical University of Civil Engineering Bucharest (Sanitary Engineering and Water Protection Department), Romania 2007. E-mail: drengabdul@hotmail.com It has been known since 1830 that milk contains two types of protein which can be separated by acidification to pH 4.6. The proteins insoluble at pH 4.6 are called caseins and represent ≈ 78% of the total nitrogen in bovine milk; the soluble proteins are called whey or serum proteins (Gerrit, 2003). Lactose is the principal carbohydrate in the milks of all mammals; non-mammalian sources are very rare. Milk contains only trace amounts of other sugars, including glucose (50 mg/l), fructose, glucosamine, etc.

1. Introduction
The dairy industry is characterized by the multitude of products and therefore production lines. Plants can have as few as one or two production lines or all of them (pasteurized milk, cheese, butter, etc.). Service and ancillary units provide water and energy requirements as well as maintenance, storage, packaging, testing and analysis needs. Because of the nature of milk and milk products, which are susceptible to microbial spoilage, equipment is characterized by designs which facilitate hygienic operation, easy cleaning and sterilization. While many older plants use open equipment and batch processing, modern dairy food plants used closed systems operated continuously for periods up to 24 hours. Shut down for cleaning is generally required at least once per day.

3. Dairy Processing
The dairy industry is divided into two main production areas: 1- The primary production of milk on farms-the keeping of cows (and other animals such as goats, sheep etc.) for the production of milk for human consumption; 2- The processing of milk with the objective of extending its saleable life. This objective is typically achieved by: (a) heat treatment to ensure that milk is safe for human consumption and has an extended keeping quality, and (b) preparing a variety of dairy products in a semidehydrated or dehydrated form (butter, hard cheese and milk powders), which can be stored. Dairy processing occurs world-wide; however the structure of the industry varies from country to country. In less developed countries, milk is generally sold directly to the public, but in major milk producing countries most milk is sold on a wholesale basis. In Ireland and Australia, for example, many of the largescale processors are owned by the farmers as cooperatives, while in the United States individual contracts are agreed between farmers and processors. Dairy processing industries in the major dairy producing countries have undergone rationalisation, with a trend towards fewer but larger plants operated by fewer people. As a result, in the United States, Europe, Australia and New Zealand most dairy processing plants are quite large. Plants producing market milk and products with short shelf life, such as yogurts, creams and soft cheeses, tend to be located on the fringe of urban centres close to consumer markets. Plants manufacturing items with longer shelf life, such as butter, milk powders, cheese and whey powders, tend to be located in rural areas closer to the milk supply.

2. Milk
Milk is the nutrient fluid secreted by the mammary glands of female mammals (including monotremes). The female ability to produce milk is one of the defining characteristics of mammals. It provides the primary source of nutrition for newborns before they are able to digest more diverse foods. The early lactation milk is known as colostrum, and carries the mother's antibodies to the baby. It can reduce the risk of many diseases in both the mother and baby. Milk from domesticated animals has been used by humans since at least 8000 BC. Although sheep and goats were the first domesticated dairy animals, because they are more easily managed than cattle, but now the cattle are the dominant dairy animals. Total recorded world milk production is ≈ 600 mil tonnes per annum, of which ≈ 85% is bovine, 11% is buffalo and 2% each is from sheep and goats (Gerrit, 2003). The proportions of total world milk production used for the principal dairy products are: liquid (beverage) milk, ≈ 39%; cheese, ≈ 33%; butter, ≈ 32%; whole milk powder, ≈6%; skimmed milk powder, ≈ 9%; concentrated milk products, ≈ 2%; fermented milk products, ≈ 2%; casein, ≈ 2%; and infant formulae, ≈ 0.3%. Milk is a very complex liquid consisting of over 100,000 different molecules. The gross composition of milk is defined as the fat, protein, lactose, ash, and total solids content. For cow's milk the gross composition is 4.1% fat; 3.6% protein; 4.9% lactose; 0.7% miscellaneous components including minerals, vitamins, and gases; and the balance in water (Hui, Y. K., 1993). The fat in milk is comprised mainly of triglycerides containing a wide range of fatty acids, which in turn contain a relatively high proportion of short-chain and saturated fatty acids.

3.1 Production of pasteurized and UHT milk
The main operations for producing the pasteurized and ultra high temperature (UHT) milks include: * Receiving and testing raw milk: Raw milk is received from the collection centers which are either privately owned or government owned (public sector).

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Milk is tested for quality by the facility lab, which measures the following parameters: fat content, solid content, presence of preservatives (formaldehyde). The price of received milk is determined according to the measured quality and consequently reduced for lower fat or protein content. Milk is rejected if formaldehyde is present. * Straining: The accepted milk is then passed through strainers then to a volume-measuring device for quantification. Milk is then cooled to 6-8 ْC and stored in storage tanks, some for cow milk and others for buffalo milk. * Pasteurization: Milk is heated in two stages, first from 7 to 65 ْC, and then from 65 to 80 ْC. It is kept at 80 ْC for 15 sec, and then cooled to 4-6 ْC. The heating and cooling processes are applied to ensure that most commonly present bacteria are killed and accordingly guarantee public safety. * UHT milk: Pasteurized milk is sterilized by raising the temperature to 135–150 ْC for 4 seconds (Ultra High Temperature, UHT). Milk is then homogenized by reducing the size of fat globules to prevent separation of cream on the surface. * Packaging: Milk is introduced in an automatic filling machine that usually used polyethylene bags.

Fig. 1 Principal stages of cheese-making process (Smith, 1995)

3.3 Dairy Processing Effluents
The volume, concentration, and composition of the effluents arising in a dairy plant are dependent on the type of product being processed, the production program, operating methods, design of the processing plant, the degree of water management being applied, and subsequently the amount of water being conserved. Dairy wastewater may be divided into three major categories: 1- Processing waters, which include water used in the cooling and heating processes. These effluents are normally free of pollutants and can with minimum treatment be reused or just discharged into the storm water system. 2- Cleaning wastewaters emanate mainly from the cleaning of equipment that has been in contact with milk or milk products, spillage of milk and milk products, whey, pressings and brines, CIP cleaning options, and waters resulting from equipment malfunctions and even operational errors. 3- Sanitary wastewater, which is normally piped directly to sewage works. Before the methods of treatment of diary processing wastewater can be appreciated, it is important to be acquainted with the various processes involved in diary product manufacturing and the pollution potential of different dairy products (Table 2). Wastewater loading for the American dairy industry is summarized in Table 3. Table 2 Reported BOD and COD values for typical dairy products (Wang & Howard, 2004) BOD5 mg/l COD mg/l Product Whole milk 114,000 183,000 Skim milk 90,000 147,000 Butter milk 61,000 134,000 Cream 400,000 750,000 Evaporated milk 271,000 378,000 Whey 42,000 65,000 Ice cream 292,000 -

3.2 Cheese
Cheese is one of mankind's oldest foodstuffs. It is nutritious. There are at least three constants in cheesemaking: milk, coagulant, and culture. By introducing heating and salting steps in cheesemaking, a potential for numerous varieties has been realized. On a global scale, 30% of all milk is used for cheese; the proportion is about 40% in North America and about 50% in the European Union (Fox. P. F, 1998). Although traditional cheeses have a rather high fat content, they are rich sources of protein and in most cases of calcium and phosphorus and have anticarigenic properties; some typical compositional data are presented in Table 1. There are at least 1000 named cheese varieties, most of which have very limited production (Fox. P. F, 1998). The principal families are Cheddar, Dutch, Swiss and Pasta filata (e.g. Mozzarella), which together account for about 80% of total cheese production.
Table 1 Composition of selected cheese per 100 g (Fox. P. F, 1998)

Cheese type Cheddar Cottage Cream cheese Feta Mozzarella

Water (g) 36 79.1 45.5 56.5 49.8

Protein (g) 25.5 13.8 3.1 15.6 25.1

Fat (g) 34.4 3.9 47.4 20.2 21

Typically, five steps, or groups of steps, are involved in the conversion of milk to cheese curd: (1) coagulation, (2) acidification, (3) syneresis (expulsion of whey), (4) molding/shaping and (5) salting The basic stages of cheese-making shown in Fig. 1.

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Table 3 American dairy and milk processing plant effluent loadings (Cast, 1995)
Products Milk Cheese Ice cream Condensed milk Powder Cottage cheese Cottage cheese and milk Cottage cheese, ice cream, and milk Mixed products Wastewater (kg ww/kg milk) range 0.10-5.40 1.63-5.70 0.80-5.60 1.00-3.30 1.50-5.90 0.80-12.40 0.05-7.20 1.40-3.90 0.80-4.60

Milk has BOD content 250 times greater than that of sewage. It can therefore be expected that dairy wastewaters will have relatively high organic loads, with the main contributors being lactose, fats, and proteins (mainly casein), as well as high levels of nitrogen and phosphorus that are largely associated with milk proteins. The COD and BOD for whey have, for instance, been established to be between 35,000-68,000 mg/l, with lactose being responsible for 90% of the COD and BOD contribution (Wang & Howard, 2004).

4. Treatment of Dairy Wastewater
4.1 Introduction
The dairy industry is generally considered to be the largest source of food processing wastewater in many countries. As awareness of the importance of improved standards of wastewater treatment grows, process requirements have become increasingly stringent. Although the dairy industry is not commonly associated with severe environmental problems, it must continually consider its environmental impact, particularly as dairy pollutants are mainly of organic origin. For dairy companies with good effluent management systems in place, treatment is not a major problem, but when accidents happen, the resulting publicity can be embarrassing and very costly. All steps in the dairy chain, including production, processing, packaging, transportation, storage, distribution, and marketing, impact the environment. Owing to the highly diversified nature of this industry, various product processing, handling, and packaging operations create wastes of different quality and quantity, which, if not treated, could lead to increased disposal and severe pollution problems. In general, wastes from the dairy processing industry contain high concentrations of organic material such as proteins, carbohydrates, and lipids, high concentrations of suspended solids, or high biological oxygen demand (BOD) and chemical oxygen demand (COD), high nitrogen concentrations, high suspended oil and/or grease contents, and large variations in pH, which necessitates "specialty" treatment so as to prevent or minimize environmental problems. The dairy waste streams are also characterized by wide fluctuations in flow rates, which are related to discontinuity in the production cycles of the different products. These aspects increase the complexity of the wastewater treatment process.

The problem for most dairy plants is that waste treatment is perceived to be a necessary evil; it ties up valuable capital, which could be better utilized for core business activity. Dairy wastewater disposal usually results in one of three problems: (a) high treatment levies being charged by local authorities for industrial wastewater; (b) pollution might be caused when untreated wastewater is either discharged into the environment or used directly as irrigation water; and (c) dairy plants that have already installed an aerobic biological system are faced with the problem of sludge disposal. To enable the dairy industry to contribute to water conservation, an efficient and cost-effective wastewater treatment technology is critical. The level of treatment is normally dictated by environmental regulations applicable to the specific area. While most of larger dairy factories have installed treatment plants or, if available, dispose of their wastewater into municipal sewers, cases of wastewater disposal into the sea or disposal by means of land irrigation do occur. In contrast, most of smaller dairy factories dispose of their wastewater by irrigation onto lands or pastures. Because the dairy industry is a major user and generator of water, it is a candidate for wastewater reuse. Even if the purified wastewater is initially not reused, the dairy industry will still benefit from in-house wastewater treatment management, because reducing waste at the source can only help in reducing costs or improving the performance of any downstream treatment facility. Best practice of diary wastewater treatment is given in Fig. 2. Segregation Screening Equalisation
pH Control Fat Removal BOD Removal

Land Irrigation Fig. 2 Best practice for dairy wastewater treatment

4.2 Treatment Options
The highly variable nature of dairy wastewaters in terms of volumes and flow rates (which is dependent on the factory size and operation shifts) and in terms of pH and suspended solid (SS) content (mainly the result of the choice of cleaning strategy employed) makes the choice of an effective wastewater treatment regime difficult. Because dairy wastewaters are highly biodegradable, they can be effectively treated with biological wastewater treatment systems, but can pose a potential environmental hazard if not treated properly.

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The three main options for the dairy industry are: (a) discharge to and subsequent treatment of factory wastewater at a nearby sewage treatment plant; (b) removal of semisolid and special wastes from the site by waste disposal contractors; or (c) the treatment of factory wastewater in an onsite wastewater treatment plant. According to Robinson (1994), the first two options are continuously impacted by increasing costs, while the control of allowable levels of SS, BOD, and COD in discharged wastewaters are also becoming more stringent.

1- Hydrolysis 2- Acidogenesis & Acetogenesis 3- Methanogenesis
* Stage -1- Hydrolysis

Since bacteria are unable to take up particulate organic matter, the first step in anaerobic degradation consists of the hydrolysis of polymers through the action of exo-enzymes to produce smaller molecules which can cross the cell barrier. Two types of enzymes are involved in substrate degradation: Endoenzymes and exoenzymes as shown in Fig. 3.

4.3 Biological Treatment
Biological degradation is one of the most promising options for the removal of organic material from dairy wastewaters. However, sludge formed, especially during the aerobic biodegradation processes, may lead to serious and costly disposal problems. This can be aggravated by the ability of sludge to adsorb specific organic compounds and even toxic heavy metals. However, biological systems have the advantage of microbial transformations of complex organics and possible adsorption of heavy metals by suitable microbes. Biological processes are still fairly unsophisticated and have great potential for combining various types of biological schemes for selective component removal (Wang & Howard, 2004). 4.3.1 Aerobic Treatment Aerobic biological treatment methods depend on microorganisms grown in an oxygen-rich environment to oxidize organics to carbon dioxide, water, and cellular material. Systems of aerobic treatment can include the conventional activated sludge process, the rotating biological contactors, the conventional trickling filters, etc. 4.3.1 Anaerobic Treatment Anaerobic treatment is a biological process ideally suited for the pretreatment of high strength wastewaters that are typical of many industrial facilities today. Anaerobic digestion is a process by which microbes are used in the absence of oxygen for the stabilization of organic matters by conversion to biogas (methane and carbon dioxide), new biomass and inorganic products. Up to 95% of the organic load in a waste stream can be converted to biogas (methane and carbon dioxide) and the remainder is utilized for cell growth and maintenance. The process reactors are covered to prevent the introduction of air and the release of odor.
4.3.1.1 Anaerobic metabolism and biochemical pathways

Fig. 3 Types of enzymes (Michael, 2003) All bacteria produce endoenzymes, but not all bacteria produce exoenzymes. Each exoenzyme as well as each endoenzyme degrades only a specific substrate or group of substrates. Therefore, a large and diverse community of bacteria is needed to ensure that the proper types of exoenzymes and endoenzymes are available for degradation of the substrates present. The relative abundance of bacteria within an anaerobic digester often is greater than 10 cells per millilitre (Michael, 2003). The large insoluble molecules consisting of many small molecules joined together by unique chemical bonds. The small molecules are soluble and quickly go into solution once the chemical bonds are broken. Hydrolytic bacteria or facultative anaerobes and anaerobes that are capable of performing hydrolysis achieve breakage of these unique bonds. Hydrolysis is in most cases, notably with sewage as substrate, rate-limiting for the overall process of anaerobic degradation of organic matter and is very sensitive to temperature. For that reason, design of the anaerobic reactors for sewage treatment is usually based on the hydrolysis step. Anaerobic digesters at industrial wastewater treatment plants that degrade simplistic, soluble organic compounds such as glucose do not experience hydrolysis stage.
* Stage -2- Acidogenesis & Acetogenesis
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The biochemistry of anaerobic organisms is quite similar to aerobic organisms to some extent. In case of anaerobic organisms, oxygen can not serve as the electron acceptors. In fermentation, the electron donors are usually organic compounds. Generally, anaerobic bacteria may be divided into two groups: 1) facultative anaerobes and 2) anaerobes. The anaerobic digestion process and production of methane is divided into stages. Three stages often are used to explain the sequence of microbial events that occur during the digestion process and the production of methane. These stages are:

During the acidogenesis (or fermentation process), the hydrolysis products which are relatively small, soluble compounds are diffused inside the cells of facultative anaerobes and anaerobes through the cell membrane and then are either fermented or anaerobically oxidized. These processes occur by a complex consortium of hydrolytic and non-hydrolytic microorganisms which are the source of energy for the acidifying population. The degradation of these compounds results in the production of CO2, H2S, alcohols, organic acids, some organic-N compounds, and

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some organic-sulfur compounds. The most important of the acids is acetate. Acetate is the principal organic acid or volatile acid used as a substrate by methane-forming acteria. The short chain-fatty acids, other than acetate, that are produced in the acidogenesis step are further converted to acetate, hydrogen gas and carbon dioxide by the acetogenic bacteria. The available H2 and CO2 are partly converted into acetate. Both propionate and butyrate acids are important intermediates in anaerobic digestion, and then are converted by the hydrogen producing acetogenic bacteria into acetate and hydrogen. The methane-forming bacteria can not use propionate and butyrate directly to produce CH4. Acetogenic bacteria reproduce very slowly; generation time for these organisms is usually greater than 3 days.
* Stage -3- Methanogenesis

The reproductive times or generation times for methane-forming bacteria range from 3 days at 35°C to 50 days at 10°C. Because of the long generation time of methane-forming bacteria, high retention times are required in an anaerobic digester to ensure the growth of a large population of methane-forming bacteria for the degradation of organic compounds. At least 12 days are required to obtain a large population of methane-forming bacteria. There are three principal groups of methaneforming bacteria: 1) the hydrogenotrophic methanogens, 2) the acetotrophic methanogens, and 3) the methylotrophic methanogens The use of different substrates by methane-forming bacteria results in different energy gains by the bacteria. For example, hydrogen-consuming methane production results in more energy gain for methane-forming bacteria than acetate degradation. Although methane production using hydrogen is the more effective process of energy capture by methane-forming bacteria, less than 30% of the methane produced in an anaerobic digester is by this method. Approximately 70% of the methane produced in an anaerobic digester is derived from acetate. There are many factors affecting performance of anaerobic processes such as: pH, temperature, alkalinity, nutrients, toxic substances and retention times. High-strength wastes are usually treated in anaerobic suspended growth systems, whereas soluble wastewaters are usually treated in anaerobic fixed-film systems. Table 4 shows a summary of efficiencies of some biological treatment systems used to treat dairy effluents.

In the methanogenic stage, methane is formed mostly from acetate and both carbon dioxide and hydrogen gas. Methane is also formed from some organic compounds other than acetate. Therefore, all other fermentative products must be converted to compounds that can be used directly or indirectly by methane-forming bacteria. Acids, alcohols, and organic-nitrogen compounds that are not degraded by methane-forming bacteria accumulate in the digester supernatant. Methane-forming bacteria are oxygen-sensitive, fastidious anaerobes and are free-living terrestrial and aquatic organisms. All methane-forming bacteria produce methane. No other organism produces methane. Methane-forming bacteria obtain energy by reducing simplistic compounds or substrates like CO2 and acetate (CH3COOH). Some methane-forming bacteria are capable of fixing molecular nitrogen (N2).

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5. Anaerobic Treatment of Whey in Stirred Batch Reactor (AnSBR)
This research orientation was the biological treatment of whey using anaerobic process. The study was carried out in laboratory scale with anaerobic stirred batch reactor, using fresh whey. The main focuses were investigating the feasibility of a biological process for treating highly concentrated carbohydrate-containing wastewater under anaerobic conditions and also the monitoring of biogas production and COD reduction. Anaerobic operation has recently been accepted as an effective mean of treating high strength wastewaters. For wastewater with a total BOD in excess of 4000 mg/1, anaerobic contact process was cheaper than an aerobic process. There were many applications which were described for wastewater from the meat packing, brewing, pharmaceutical and food processing industries. There are some benefits of anaerobic processes in treating organic wastewater, which can be summarized as follows: (1) Low cellular yields. This process requires only relatively few inorganic nutrients for treating wastewater and produces only little amounts of surplus sludge. (2) The major gas product is methane, a valuable and sustainable energy resource. The second major gas is carbon dioxide. (3) No requirement of oxygen in the process, in contrary strictly anaerobic conditions and the absence of nitrate are required. (4) Good capability for degrading a variety of natural organic compounds. Produce a residual sludge which is inoffensive and useful as a soil conditioner and low grade fertilizer. (5) Anaerobic digestion of primary sewage sludge effectively reduces the concentration of pathogenic microorganisms.

5.2 Experiment Design
5.2.1 Laboratory-scale anaerobic stirred batch reactor The reactor used for the study was glass vessel with a theoretical volume of 2.5 L, and 2 litres of working volume. The reactor had an off-gas line attached to the lid of the vessel. A scheme of the stirred batch reactor, which was used in this study, is shown in Fig. 4. The primary components of the system were the main reactor compartment, the wastewater supplying system, thermometer, the constant temperature jacket providing by automatic pumping of hot air, pH electrode, pH adjustment system, magnetic stirrer device and the biogas measuring system. The reactor was surrounded by a hot-air-jacketed glass vessel. Fresh whey (wastewater) feed was added into the system in different concentrations. Biogas left the reactor through a port at the top and was measured by water displacement. Hot air was pumped through the air-jacket surrounding the reactor to maintain the temperature at 38 ْC.

Fig. 4 A Scheme of anaerobic stirred batch reactor
Where: 1. Reactor, 2. Hot-air Jacket, 3. Air heating device, 4. pH electrode, 5. Thermometer, 6. Buffering solution tank, 7. Wastewater (whey) feed tank, 8. Digital reading device of pH and temperature, 9. Biogas pipe, 10. Releasing biogas pipe to the air, 11. Samples taking pipe, 12. Discharge tap, 13. Biogas measuring apparatus, 14. Magnetic stirrer device, 15. Automatic control device of hot air pumping, 16. Electricity source.

5.1 Whey
Whey is the serum of milk resulting after removal of fat and casein. Whey is a by product in cheese and casein production. There are two kinds of whey, depending on the type of milk coagulation used, either sweet or sour whey. Sweet whey is obtained if milk is coagulated by proteolytic enzymes, such as chymosine and pepsine or microbial enzymes produced from Mucor miehei and Mucor pusillus. It comes from the production of cheddar, Swiss and Italian varieties of cheese. Acid whey is obtained if milk is coagulated by acids mainly by lactate from lactic acid fermentation of lactose. It comes from soft cheese production, with cottage cheese as a predominant product. The chemical composition of whey depends upon the type of coagulation used. It contains roughly half the solid of the whole milk from which it is derived and most of the water soluble vitamins and minerals. Typical whey contains around 6.5% total solids. These include lactose (68-72%), protein (12-13%), minerals (8-9%) plus small amounts of fat and lactic acid. Generally, 100 L of milk produces about 12 kg of cheese or about 3 kg of casein. In either case, about 87 L of whey is made as a byproduct (Marisa, 2004). 5.2.2 Reactor operation The AnSBR reactor was fed with a wastewater, in different concentrations of COD, which was prepared by diluting the fresh whey (75000 mg/l of COD), provided by a cheese maker, with water. It was anticipated that by seeding the pilot-scale digester with anaerobic sludge taken from a septic tank to reduce the time required for start-up period. The sludge volume was 25% of the wastewater used in the test (1:4 seed/feed ratio). This reactor was operated under various organic loading rates (OLR) and various hydraulic retention times (HRT). Firstly, reactor was fed with 100 ml of fresh whey per liter of water (10 % in concentration) with different retention times. The working volume of reactor (2 litres) was operated under a draw/feed regime (i.e. 200 ml of the reactor contents were drawn from the reactor prior to feeding with an equivalent volume of fresh feed). Different retention times (e.g. 1, 2, etc, of days) used in this stage. Secondly, feeding percentage (e.g. 80, 100, 200 ml, etc, of whey per litre of water) was increased step by step. The reactor performance was monitored by daily measurement of the pH, temperature and the total biogas production.

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The COD for both influent and effluent and the biogas production were measured. The pH in the reactor was measured automatically by a pH electrode unit. pH controlling is very important, especially to the methanogenic stage. The pH adjustment process accomplished manually by adding buffering capacity, i.e alkaline solutions. Buffering capacity can be increased with addition of bicarbonates (e.g. NaHCO3) or compounds reacting with carbon dioxide and thus producing bicarbonates (e.g. NaOH). The pH value was kept in range 6.5 - 7.5 (Shanta, 2004). In certain cases, the pH adjusting and the temperature controlling were stopped in order to determine the reactor capabilities for degrading organic pollutants. Fig. 5 is shown the pilotscale used in the experiment. Finally, the reactor performance was monitored after adding minerals (e.g. FeC13).

100 90 Organic removal % 80 70 60 50 40 30 20 10 0 0 10 20 30 40 Organic loading rate (OLR), (g COD/l.d)

Fig. 7 Organic removal efficiency at different OLR The production of biogas, in our study, increased continuously, day by day, until reaching to a constant value nearly. The maximum production of biogas was 0.32 litre per gram of COD removed (or nearly 20.8 L of biogas per litre of whey) as shown in Fig. 8.
Biogas production (l/g COD removal) 0.35 0.3 0.25 0.2 0.15 0.1 0.05 0 0 5 10 15 20 25 30 35 Time of test running (days)

Fig. 5 A Pilot-scale of anaerobic stirred batch reactor

Fig. 8 The maximum biogas production per g CODremoved Ferric ions addition has an improving effect on methanogenesis by increasing the methane production and acetate conversion rate. The methane production increases higher than theoretically expected by adding ferric ions. The additional of ferric ions improve methanogenesis. It increases the methane production during the exponential phase. By adding 50 mg/l of FeC13 to the AnSBR in our experiment, the biogas production increased nearly 50% as shown in Fig. 9.

5.3 Results
The AnSBR reactor was fed with fresh whey (75000 mg of COD per litre) diluted with water in different concentrations. The start up stage was used 10 % of diluted whey (100 ml of whey per litre of water or 7.5 g COD/l). The AnSBR mixed completely by magnetic stirrer (200-400 rpm). After many weeks of start-up operation, a constant concentration of organic feeding (10 g COD/l.d) was applied at various hydraulic retention times. The organic removal efficiencies are shown in Fig. 6. A COD removal efficiency of >= 76 % was achieved when the reactor was operated at a HRT longer than 5 days and an OLR lower than 10 g COD/l.d (Figure 6 and 7). The COD removal efficiency decreased to less than 74 % when the OLR was increased above 15 g COD/l.d. The maximum removal of COD was 87% for 7.5 g COD/l.d of OLR.
100 90 Organic removals % 80 70 60 50 40 30 20 10 0 0 5 10 HRT (days) 15 20

Fig. 9 Effect of FeC13 adding to the AnSBR on biogas production. (A) Without adding FeC13, (B) With adding 50 mg/l of FeC13. Finally, the current study of using AnSBR as a pilot scale is a modest step to explain the capability of the anaerobic stirred batch reactor to be a primary stage of treatment of high organic wastewater. Interesting results have made, but more investigations should be achieved.

Fig. 6 Organic removal efficiency at different HRT

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