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Available online at www.sciencedirect.com Bioresource Technology 100 (2009) 50–58 Aerobic selectors in slaughterhouse

Available online at www.sciencedirect.com

Available online at www.sciencedirect.com Bioresource Technology 100 (2009) 50–58 Aerobic selectors in slaughterhouse

Bioresource Technology 100 (2009) 50–58

Bioresource Technology 100 (2009) 50–58 Aerobic selectors in slaughterhouse activated sludge

Aerobic selectors in slaughterhouse activated sludge systems: A preliminary investigation

Nayef Z. Al-Mutairi *

Civil Engineering Department, Kuwait University, P.O. Box 5969, Safat 13060, Kuwait

Received 25 September 2006; received in revised form 2 December 2007; accepted 3 December 2007 Available online 15 July 2008


Filamentous bulking at a slaughterhouse activated sludge treatment plant significantly reduced mixed liquor settling properties, which caused many operational problems and worsening in effluent quality. The main cause of this condition was attributed to significant levels of influent readily biodegradable COD, which was present primarily in the form of organic acids. An aerobic selector was chosen to eradicate the usual bulking incidents of slaughterhouse wastewater treatment plants. Other plant enhancements included increased aer- ation batch reactor volume, and provision of step feed capability. Comparison of data before and after aerobic selector installation showed a significant improvement in mixed liquor settleability, which eradicated the need for chemicals that had been used to control filaments and to control effluent solids loss. The additional volume of the aeration and chemicals eliminations from the activated sludge system also served to eliminate aquatic toxicity in the treated effluent. 2007 Elsevier Ltd. All rights reserved.

Keywords: Slaughterhouse wastewater; Selector; Filamentous bulking; Volatile fatty acids; Toxicity

1. Introduction

The activated sludge process has found vast application as an efficient means of treating wastewater. The purpose of the activated sludge process is to remove soluble and insoluble organics from the wastewater and to change this material into a flocculent microbial suspension that sinks well in a conventional tank. In most cases, the nature of the wastewater will impose the preferred process modifica- tions, mainly for the purpose of maintaining appropriate mixed liquor settling properties ( Eckenfelder and Mus- terman, 1995 ). Basically, activated sludge comprises a microbiological enrichment culture consisting of a mixed, and largely uncontrolled, consortium of micro- and macro-organisms ( Richard, 1989 ). It is important to maintain the growth of floc-forming bacteria on wastewater organics, which will

* Tel.: +965 4817524. E-mail address: nayef@civil.kuniv.edu.kw

0960-8524/$ - see front matter 2007 Elsevier Ltd. All rights reserved.


settle easily under gravity in the final clarifier to obtain a clarified supernatant and a thickened return sludge. How- ever, not all bacteria in the activated sludge process are floc-formers. Many different types of filamentous bacteria have been identified in activated sludge and play important roles in wastewater treatment. Filamentous bacteria directly affect sludge settling as they make provision for the rigid support network or backbone upon which floc- forming bacteria can adhere and grow into suitable acti- vated sludge flocs ( Richard, 1989 ). Filamentous bacteria may be considered detrimental to wastewater treatment when they occur in excessive quantities, but are just as important in the development of activated sludge flocs with proper settling and clarification properties (Gerardimh et al., 1990 ). Filamentous organisms grow out of the activated sludge flocs and lead to a diffuse and very irregular shape of the activated sludge flocs. This phenomenon can be either clas- sified as bulking sludge, floating sludge or foam. Bulking sludge is defined as sludge with a sludge volume index

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(SVI) of more than 150 mL/g. Floating sludge or foam builds a stable layer at the surface of the aeration basin or final clarification basin. Filamentous microorganisms can also be good indicators of conditions prevailing in an activated sludge system on a microbiological level. Low organic sludge loads (<0.1 kg BOD 5 /(kgTSS d)), low wastewater temperatures (12–15 C) and high sludge ages are considered favoring factors to the abundant develop- ment of Microthrix parvicella , which is the most frequently occurring organism ( Knoop and Kunst, 1998 ). The indica- tions given by the filamentous bacteria could also be of low dissolved oxygen (DO) (e.g. Sphaerotilus natans ), presence of septic waste (e.g. Thiothrix spp.), nutrient deficiency (e.g. Haliscomenobacter hydrossis ) and low pH in the system (e.g. fungi) (Jenkins et al., 1993 ). Furthermore, the existence of volatile fatty acids (VFAs) in wastewater has long been linked with filamentous phe- nomena (Richard and Kampfer, 1997 ). VFAs are the pri- mary constituent of readily biodegradable COD in influent slaughterhouse wastewaters. Direct measurements of changes in specific organic pools in slaughterhouse wastewater have not been performed so far. Richard and Kampfer (1997) measured protein, carbohydrate, and VFAs in two types of wastewater of different ages and found significant differences in composition. Unfortu- nately, little research has focused on the production of VFAs from slaughterhouse wastewater. In addition, practi- cal design criteria for the purification process specifically oriented toward slaughterhouse WWTPs are not generally available. Since the introduction of conventional activated sludge systems, sludge bulking has been one of the major prob- lems affecting every biological process (Bitton, 1999 ). Two toxicants, chlorine and hydrogen peroxide, have been used to selectively kill filamentous organisms in conven- tional activated sludge systems and therefore alleviate symptoms of activated sludge bulking ( Jenkins et al., 1993 ). Chlorination is however, a last alternative in the control of bulking. It should only be contemplated when effluent from the bulking plant is likely to cause environ- mental damage to the receiving water. If used correctly it provides a rapid alleviation of bulking, but if used incor- rectly, it may completely inhibit all treatment (Horan, 1990 ). On the other hand, Hwang and Tanaka (1998) showed that the application of chlorination had no effect on M. parvicella reduction or foaming suppression and that a more effective and economical method for the control thereof still needs to be established. Often, non-specific methods are used to combat bulking or floating sludge. The use of alum and iron salts or the application of strong oxidizing agents belongs to these methods. One disadvantage of using chemicals is that floc-forming bacteria are also influenced negatively. This can lead to the failure of the treatment process. Addition- ally, the use of chemicals results in an increase in toxicity and in sludge mass, which has to be disposed of or recycled ( Al-Mutairi, 2006 ).

Theoretically, the massive growth of filamentous organ- isms could be avoided by increasing the sludge load, but a higher sludge load leads to a decrease of the sludge age. The cause for the excessive growth of filamentous organ- isms can be controlled by specific measures. A more detailed understanding of the physiology and biochemistry of filamentous bacteria is still required for effective long- term control of bulking and foaming. For example, by improving wastewater quality (e.g. the prevention of H 2 S formation) or by applying modifications in the operation mode (e.g. aerobic or anaerobic selector) filamentous organisms are taken out of the system. Moreover, plant configuration provides operational conditions that sup- press filamentous bacteria and favor growth of floc-form- ers, leads to high sludge settleability characteristics (Vaiopoulou et al., 2007 ). Mueller et al. (2000) successfully tested a side-by-side full-scale aeration of a modified con- tact stabilization process incorporating an anaerobic selec- tor to control filamentous bacteria in wastewater. The application of selector reactor technology has become the promoted method for control of filament pro- liferation to enhance sludge settleability in activated sludge systems (Pujol and Canler, 1994; Kruit et al., 1993; Eikel- boom, 1994 ). A successful application of selectors relies on detailed knowledge of: (a) physiology and substrate requirement of the filamentous microorganisms, (b) waste- water composition and (c) substrate removal kinetic in the selector system ( Andreasen et al., 1999 ). Madoni and Davoli (1997) noticed a complete suppression of the growth of nocardio forms was obtained where returned activated sludge and wastewater were mixed together under both anoxic conditions and high sludge loading. However, under moderate sludge loading, the selector suppressed the growth of nocardio forms but was not able to improve set- tlement of the sludge and prevent the growth of other fila- mentous microorganisms responsible for foaming, such as M. parvicella. A principal components analysis of patterns and level of microbial activity suggested that microbial communities statistically differentiate between the selector and conven- tional activated sludge system. There was seasonal varia- tion in the structure and function of the microbial community in conventional samples while, for the selector system, there were no identifiable differences between the data communities ( Al-Mutairi, 2007 ). Aerobic selectors are an engineered system that uses dif- ferential growth kinetics to promote the development of floc-forming bacteria rather than filamentous bacteria. This is accomplished because most floc-forming bacteria grow faster than filamentous bacteria at higher BOD load- ing rates. Conversely, most filamentous organisms grow faster than floc-forming organisms at lower BOD loading rates. Basically, the process goal is to assimilate as much of the soluble organics as fast as possible, thereby denying the filamentous microbes this food source. At high BOD concentrations, the floc-forming bacteria have a higher BOD uptake rate than the filamentous bacteria and so


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the floc-forming bacteria are more competitive for the food source. This action enhances the growth and formation of better settling, floc-forming microbes, and limits the growth rate of the filamentous organisms. Furthermore, biological treatment of slaughterhouse wastewaters may not be straightforward due to high varia- tions in flow and chemical characteristics. Those factors, combined with low temperatures in the months of Decem- ber–February and high temperatures in the months of May–August may make consistent biological treatment difficult ( Al-Mutairi et al., 2003 ). The selector is also employed to smooth out fluctuations in influent composi- tion and allowing the wastewater treatment plant to run more uniformly. Influent wastewater characteristics can be impacted by fluctuations in temperature and pH, as well as flow surges. Proper design of the selector includes pro- viding the correct retention time under aeration, with the flexibility to change this as wastewater characteristics change. Adjustable dissolved oxygen targets, avoidance of prolonged over-aeration, variable return activated sludge (RAS) rates, control of final tank blanket levels, etc. are all useful tools to reduce the effects of foaming and bulking. Low F/M bulking ( Microthrix parvicella , type 0092, type 0041 and Nostocida I ) and foaming (Microthrix parvi- cella , and Nocardia sp.) can be controlled in a selector by inducing a high substrate gradient in the selector zone which stimulates floc forming microorganisms with high substrate uptake and storage rates which allows them to out compete over the filamentous microorganisms through the resulting feed–starve cycle. The experimental plan was divided into two parts. In the first part, experiments were conducted using a conventional activated sludge unit for two years and in the second, using a contact/aeration unit (selector) for another year. The objective of the present work was to compare the selector process with the conventional activated sludge process in terms of COD treatment capacity, oxygen uptake rate and toxicity. In addition, assess the selector process perfor- mance in terms of process capability to enhance final clar- ification and settleability. Furthermore, the study results are expected to be used to adjust effluent treatment technol- ogies or regulations, so that the potential threat to the envi- ronment by such materials will be reduced.

2. Plant description

Hawally wastewater treatment plant (WWTP) used for this study, a relatively small plant ( 360 m 3 /day), was designed to use activated sludge process to achieve second- ary treatment. The existing biological reactors are config- ured as a complete mix system and are limited to MLSS concentrations of 1800–2000 mg/L because of the deleteri- ous effects of sludge bulking and foaming at higher concen- trations. This system was prone to prolonged bulking periods, with Thiothrix I and II, Nostacoida limicola, S. natans , H. hydrossis , and Types 0411, 1701, 0041, 0803,

0914, and 0961 being identified in the mixed liquors. In light of the new permit limits, the WWTP’s desire to expand the facility by 25%, and given the extreme space limitations on the existing site, increased process control is required to allow higher operating MLSS concentrations to achieve the desired treatment. Furthermore, Nakhla and Lugowski (2003) tested vari- ous measures at a food-processing wastewater treatment plant to prevent sludge bulking by type 0041 and 0675 fil- amentous microorganisms. The aerobic selector was the most successful system used, reducing SVIs to 79 cm 3 g 1 in 2 weeks. Sludge settleablity was found to be inversely proportional to the aerobic selector food-to-microorgan- ism ratio. The optimal aerobic selector loading was found to be 1.8–2.7 kg BOD 5 kg MLVSS d 1 . Hawally WWTP was evaluated to define potential oper- ational changes and upgrades that would allow achieve- ment of the permit limits consistently. Finally, the WWTP was upgraded with an aerobic biological selector to improve sludge settling characteristics. The primary treatment in Hawally WWTP is achieved by a rotary screen and a dissolved air flotation unit (DAF). Fat, oil, grease (FOG), settleable solids, and sand in the ‘slaughterfloor’ effluent are removed in the flotation unit. The pretreated water will flow to the aeration tank of the biological treat- ment system. After settling, the supernatant is decanted from the tank. A commercial flocculant known as Praestol is dosed in the feed pipe to the second stage flotation unit to concentrate the colloidal and suspended pollutants in a floc-like agglomerate which can be separated from the water by flotation. Flocs, solids, and FOG are removed by the second stage DAF process. The WWTP’s original design-rated monthly average capacity: flow – 310 m 3 /day and COD – 1962 kg/day. The original design predicted 95% BOD and TSS removal efficiencies with resultant efflu- ent concentrations of less than 30 mg/L. However, the treatment system experienced frequent equipment and pro- cess limitations and was unable to meet its design goals.

3. Methods

Crude and settled sewage of the conventional activated sludge plant and the contact stabilization plant were exam- ined. Two hundred fifty ml sterile bottle samples were used ( 3) of mixed liquor, obtained from the aerobic zones of the aeration and the selector tank. The mixed liquor sam- ples were stored half-full in the bottles so as to maintain aerobic conditions for filamentous survival during sample transit. The procedures employed for identification of filaments were those used by Jenkins et al. (1993) . Samples were evaluated microscopically ( 1000, oil immersion) so as to determine the effects of the operation of the con- ventional activated sludge systems and a selector system on the integrity of the filaments and floc structure. Respi- ration measurements were generally obtained under peri- ods of load. The VFAs (formate, acetate, propionate, n -butyrate, and isobutyrate) were measured on a Dionex

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ion-chromatograph with a suppressed conductivity detec- tor. The VFA concentrations lower than 1 mg/L could be detected. The sludge index was also observed during the time of the experiment. Other parameters were measured according to Standard Methods for the Examination of Water and Wastewater (1995) . Finally, acute bioassay is a widely used tool for adding toxicity data to biological/chemical/physical field surveys of wastewater effluents. Mean 15-min EC 50 values (a dose that produces a 50% decrease in a sublethal response) for the wastewater effluent before and after the selector instal- lation. The Microtox assay is based on the inhibition of light production of the bioluminescent microbe, Vibrio fisc- heri , in the presence of toxicants ( Microbics, 1992 ). A pho- tometer is used to provide temperature control and measurement of light emission. An M500 Microtox TM sys- tem as well as reagents and protocols followed were obtained from Microbics Corporation ( Microbics, 1992 ). Data reduction and statistical analysis were performed by Microbics statistical software.

4. Results and discussion

4.1. Analysis of bulking problems

A microbiological assessment of the slaughterhouse wastewater before the upgrade of the plant showed that many filamentous bacteria were present in the mixed liquor including Thiothrix I and II (predominant), N. limicola , S. natans , H. hydrossis , and Types 0411, 1701, 0041, 0803, 0914, and 0961. The Thiothrix spp. was so concentrated that the bacteria imparted a bright orange color to the mixed liquor. Consequently, the use of chemicals to limit bulking was followed in concomitantly with more vital corrective mea- sures. Once control is obtained, sufficient time was allowed for changes in the H 2 O 2 dose to take effect. With lower doses of H 2 O 2 (e.g., <100 mg/L), several days may pass before the effects are evidenced in the activated sludge floc. Furthermore, related operating data and wastewater parameters should be collected before, during and after the trial to assist in interpreting the results. On the other hand, filamentous bulking in Hawalli WWTPs created a sludge that was light, had a high sludge volume index, and was being washed out of the secondary clarifiers even with very high polymer dosing rates. Solids that were washed out were captured in part by the tertiary DAF clarifier. However, the recovered sludge could not ini- tially be returned to the aeration tanks, so any biomass that washed out of the secondary clarifiers was lost from the activated sludge process. As a result, the F/M increased and new high F/M filaments was established. The returned activated sludge was dosed with hydrogen peroxide, which was only partially successful in killing fila- ments. Higher doses resulted in creating conditions that were favorable to a filament (Type 0803) that is normally only found in oxygen activated sludge systems. Polymers

were added at high doses to keep effluent TSS under con- trol throughout the two years of operation (Al-Mutairi et al., 2004 ). Hydrogen peroxide was used to correct a seri- ous filamentous bulking situation or, preferably, to prevent one from occurring until adjustments can be made to remove the cause. When applied to the return activated sludge, H 2 O 2 supplies dissolved oxygen which helps restore the microbial activity necessary for effective operation, while selectively oxidizing the filaments which retard settling. Furthermore, healthy activated sludge systems normally contain about 15% polysaccharides on a dry weight basis. Hawally biomass before the installation of the selector con- tained polysaccharide concentrations as high as 60%, because of the cells’ inability to metabolize high concentra- tions of readily biodegradable COD (RBCOD) (present as VFAs in the influent). This condition is known as zoogleal bulking and is induced by inadequate availability of nutri- ents. In conditions where high RBCOD levels are present, cells respond by trying to metabolize the substrate at high rates and, in order to do so, require high concentrations of biologically available nutrients and oxygen. In this case, the biomass retain a gelatinous texture, which impeded mixed liquor settleability, elevated SVI and effluent TSS concentrations, caused foaming on the aeration tanks and a thick floating scum layer in the secondary clarifiers that was very difficult to break up and remove. RAS chlo- rination did not correct this condition either. The high doses of cationic and anionic polymers that were used to trap solids and make them heavy enough to settle, at times created a sticky sludge that the suction sludge withdrawal mechanism was unable to remove. High doses of defoamers were also used to contain the foaming in the aeration basins. The high rates of chemicals used to keep the slaughterhouse WWTP in compliance drove up operational costs extensively. Fermentation of starch in the wastewater to VFAs also caused a reduction in pH in the equalization tank. VFA levels of up to 400 mg/L as measured by distillation, were observed in the tank. Table 1 , summarizes limited VFA speciation data during both the summer and winter months. It is significant that VFAs production at elevated temperatures during summer months is nearly double that observed at lower temperatures during winter months. Consumption of readily fermented substrate may have occurred during winter months causing lower concentra- tion of VFAs.

Table 1 Speciation of VFAs in the equalization effluent

VFA species

Concentration (mg/l)

Winter months

Summer months


91 ± 22 6 ± 2 5 ± 1 3 ± 0.3 0.6 ± 0.1

166 ± 32 29 ± 5 10 ± 2 8 ± 1 7 ± 0.1






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Sodium hydroxide was added to the raw wastewater to control the septic conditions before it was pumped to the treatment system to suppress the growth of filamentous bacteria and to retain H 2 S in the dissolved form until it could be oxidized in the aeration tanks. This procedure was partially successful, but was discontinued because of its high cost and because of the heavy scale deposition in the equalization effluent pipes resulting from the change in wastewater chemistry. The impact of the termination of pH correction was to elevate the level of VFA produc- tion in the primary clarifier. In addition, a remarkable increase in VFA was observed in the equalization effluents, as represented by the increase from a mean value of 138 ± 62 mg/L in the equalization influent to 217 ± 119 mg/L in equalization effluent and then to 351 ± 112 mg/L in the equalization effluent after pH termination. One-way analysis of variance within the three means of VFA concentrations of the equalization tank showed that the difference between them were statisti- cally significant at a = 0.05 ( F = 307 and p -value = 0.00). The installation of biological selector at other treatment plant facilities showed promise for creating conditions that would favor the growth of floc forming bacteria over fila- mentous bacteria in wastewater containing high RBCOD levels (Marshall and Richard, 1999 ). However, there was little information in the literature that related directly to slaughterhouse wastewater, and the slaughterhouse man- agement had mixed success with the conventional activated sludge technology. The existing aeration basins of the Hawally slaughterhouse had not prevented the proliferation of the filamentous bacteria because they had not created a properly sized selector zone, and so they were removed. It was also evident that the existing aeration system was unable to maintain a steady acceptable DO concentration at the influent end of the aeration tanks. Also, excess air entrainment in the mixed liquor contributed to settling problems in the clarifiers. A temporary step feed system was installed to distribute the loading along the length of the aeration basins. This improved system operability, lent credibility to the concept of adopting step feed as an alter- native operating strategy.

4.2. Aerobic selector performance

The method selected to treat slaughterhouse wastewater is influenced by its characteristics. The slaughterhouse wastewater contains starch-like, low molecular weight organic compounds that are very biodegradable. Oxygen uptake rate (OUR) tests were conducted to assess the level of slaughterhouse wastewater biodegradability. Prelimin- ary OUR measurements of the influent, gave very high readings but tended towards endogenous respiration rates (in the range, 106–240 mg/g h). In addition, the results of mixed liquor respiration tests were in the range of 4– 105 mg/g h. The high values observed in the influent are likely due to the physiological stress experienced by the bacteria upon the exposure to an environment having

markedly changed conditions of temperature, dissolved oxygen and pH. The respirometry conducted, showed that the wastewa- ter RBCOD was 80–85% of the primary effluent COD. Thus, the wastewater was readily biodegradable which would, in the presence of microorganisms, exert a high demand for oxygen and nutrients. Comparable RBCOD values for sewage are 1–16% and 30–50% for other indus- trial wastewaters (Sperandio et al., 2000 ). The results of this test indicated that the slaughterhouse wastewater was amenable to treatment in an aerobic selector-assisted acti- vated sludge system. A selector is a portion of an activated sludge system that precedes the main bioreactor, receives the influent wastewater and RAS, and has a low residence time (15–30 min). This environment favors floc formers over filamentous bulking organisms, which results in mixed liquor with good settling properties. The WWTP’s upgrade design-loading monthly average capacity: flow – 650 m 3 /day and COD – 1900 kg/day. Fur- thermore, there was a concern over the frequency of peri- ods of sustained COD loadings that might not be adequately addressed by a reactor volume design based on a daily average COD loading. Review of influent COD loading data led to a maximum week condition being selected as the design basis for reactor volume. The design of aerobic selectors is controversial, with some researchers advocating multiple steps to induce an incremental F/M loading through the selector ( Jenkins

et al., 1993 ). Others base their approach on a COD floc loading ( Al-Mutairi et al., 2003 ), and others yet base their designs on complete COD uptake in the selector (Still et al.,

1996 ).

A two-cell aerobic selector was chosen over other designs to provide operational flexibility given the uncer- tainty of using selectors to treat slaughterhouse wastewa- ters. The selector volume is approximately 10% of the overall volume under aeration. The design incorporates a primary effluent bypass capability to permit diversion of excessively high loads around the first stage of the selector and to the main aeration basins. There was also a need for two new aeration tanks of the same volume as the two existing tanks to satisfy projected COD loadings. A provi- sion was made to allow step feeding of primary effluent to four locations along the length of the aeration basin. Also, a review of two years of COD loading rate data revealed periods of sustained elevated loadings that lasted as long as 6 h. This peak loading condition was used as the basis of the design of the aeration system. Following the startup of the modified treatment facility in May 2003, performance has been excellent. Fig. 1 a–d summarize monthly average effluent characteristics for 2003/2004. They show an average soluble COD removal of 79% across the selector. Soluble COD is typically slightly higher than RBCOD, which suggests that the selec- tor is functioning close to the predicted level of perfor- mance. The graphical summary of the data includes four graphs: histogram of data with an overlaid normal curve,

N.Z. Al-Mutairi / Bioresource Technology 100 (2009) 50–58


Al-Mutairi / Bioresource Technology 100 (2009) 50–58 55 Fig. 1. (a–d) Show histograms of monthly average

Fig. 1. (a–d) Show histograms of monthly average influent/effluent characteristics for 2000/2001 data with an overlaid normal curve.

boxplot, 95% confidence intervals for l , and 95% confi- dence intervals for the median. This tool, along with nor- mality assessment statistics generated using minitab univariate procedure, alerts the investigator to non-normal data distributions and the presence of outliers. The histo- gram clearly portrays information on location, spread, and shape that enables the user to perceive subtleties regarding the functioning of the chemical process that is generating the data. It can also help suggest both the nature of, and possible improvements for, the physical mechanisms at the treat- ment plant. The mean of the influent COD concentration is 1144 mg/L (95% confidence intervals of 1108 and 1182 mg/L). The standard deviation is 58 mg/L (95% con- fidence intervals of 41 and 99 mg/L). On the other hand, the mean of the selector COD concentration is 240 mg/L (95% confidence intervals of 217 and 263 mg/L). The stan- dard deviation is 36 mg/L (95% confidence intervals of 25 and 61 mg/L). The final effluent COD quality of the selec- tor is very good with a monthly average total COD ranging from 217 to 263 mg/L. A one-way analysis of variance of all the means of soluble COD showed that the difference between all the effluents were statistically significant at a = 0.05 ( F = 1075.73 and p -value = 0.00). The reliability of the selector can be assessed using prin- cipal component analysis (PCA) in terms of its ability to produce consistently acceptable reclaimed wastewater. The PCA technique is one of the most powerful and com- mon techniques used for reducing the dimensionality of large sets of data without loss of information. It implies a mathematical procedure that transforms the overall set of original variables into principal components (PCs).

The statistical results from Table 2 show, the unrotated loadings and communalities which explains 100% of the data variability and the communality values indicate that all variables are well represented by these two factors. The percent of total variability represented by the factors does not change with rotation, but after rotating, these fac- tors are more evenly balanced in the percent of variability that they represent, being 44.7% and 35.0%, respectively. Factor 1 has large negative loadings on influent COD con- centration ( 0.798), selector effluent concentration ( 0.390), and final DAF effluent ( 0.196), and a small positive loading on first DAF effluent COD concentration (0.332). Factor 2 has a large positive loading on first DAF effluent of 0.754 and small positive loadings of 0.347 and 0.434, respectively, on primary and selector effluents, and a negative loading on final DAF effluent. The first factor negatively loads on primary and on two variables, selector

Table 2 Principal component factor analysis of the correlation matrix showing the unrotated factor loadings and communalities


Factor 1

Factor 2

Factor 3 Factor 4









First DAF














Final DAF











0.4352 4.000

% Variance







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and final DAF effluents, that generally decrease when the primary effluent is reduced. We might consider factor 1 to be ‘‘primary, selector, and final DAF effluentsfactors. The second factor might be considered a ‘‘first DAF efflu- ent factor. Table 2 clearly demonstrates that, in-spite of significant variations in the quality of primary effluent, the final DAF effluent remained essentially unchanged. This indicates that the selector combined with the final DAF system played an important role in the stability of effluent quality so as to dampen variations in the quality of the effluent. The final DAF effluent consistently satisfied the water quality requirements for irrigation. Variability in the primary efflu- ent quality may be taken as an indication of an inherent diurnal or seasonal variations in influent wastewater flow and characteristics as well as process control practices. Selector operational targets vary according to change in wastewater characteristics. When the system before modifi- cation and the selector response were compared in terms of SVI, as a result of different factors impacting the plant over the past three years of operation. SVI values were highly variable before treatment system modifications. Values as high as 600 mL/g were common with peak values as high as 1100 mL/g. Furthermore, SVI significantly dropped to 250 mL/g when chlorine was added to the RAS to rapidly kill filaments rather than wait for them to die off due to changes in selector operation. However, when pH correc- tion was terminated, SVI values began to increase dramat- ically to about 900 mL/g. In addition, the installation of the selector showed an excellent SVI values in the range of 80–140 mL/g. At startup on May 2003, the selector was operated with the first cell utilized for RAS reaeration during which floc is saturated with oxygen before the influent is fed to the selec- tor, preparing the floc-forming bacteria for rapid COD uptake and utilization. When RAS and influent were both fed to the second cell, the SVI gave excellent results with SVI < 120 mL/g without RAS chlorination. However, a test of a two-cell selector operation was initiated in Septem- ber 2003 by feeding RAS and influent to the first selector cell in which the performance was not as good, with SVI in the range, 250–300 mL/g. To further test operational flexibility part of the influent was step fed around the selector in November 2003. Mini- mal changes were made to step feed flow during this period. In March 2004, the system reverted to RAS re-aeration. In June 2004, SVI values started to decline to a low of 37 mL/ g on July, 2004 and effluent turbidity increased. Step feed flow was increased to grow some filaments to serve as a floc ‘‘backbone . SVI values stabilized and slowly began to increase until August, 2004 when step feed was terminated. No step feed was used until September, 2004. The step feed flow is now adjusted based on microscopic examination of MLSS and daily SVI values. There have been times when step feed has been completely turned off but this almost always results in too few filaments to support floc develop- ment and excessive globular zooglea growth.

Table 3 Microtox TM toxicity of wastewater treatment plant effluents


EC50 (15 min test)

Activated sludge system

Selector system


15% (10–20)

13% (7–25)

Selector effluent


70% (59–85)

Final effluent

3% (3–5)


Values in parentheses are as % confidence limits calculated by Microtox TM software.

5. Toxicity assessment

Results of acute toxicity tests on whole effluents are reported as EC 50 (for effective concentration). Microtox suggested mild acute toxicity from the slaughterhouse wastewater influent. The slaughterhouse wastewater efflu- ent before the selector installation severely depressed light production in the Microtox assay. However, the selector effluent exhibited no acute toxicity (did not inhibit bacterial light production) at the highest concentrations tested (Table 3 ). The final effluent of the activated sludge system indicated an increase in toxicity from 15% to 3% of EC 50 during the tertiary chemical process, presumably due to the overdose of polymer/alum and chemicals used. How- ever, after the installation of the selector, the final effluent achieved a toxicity reduction from 13% to >100% of EC 50 . There are a number of potentially toxic constituents in the slaughterhouse effluent, but only three, alum/polymer, chlorine and ammonia, are present in sufficiently high con- centrations to produce the observed acute toxicity. Alumi- num and polymer were present at relatively high concentrations for the coagulation process and are almost certainly the chief toxicants beside ammonia ( Al-Mutairi, 2006 ). While aluminum concentrations, appear high in absolute terms (0.7–0.9 mg/L) toxicity of this metal is mit- igated at neutral pH. Although the total aluminum concen- tration appears high, it would be almost entirely particulate and would therefore have very low biological activity. Given the importance of alkalinity and hardness to toxicity of many contaminants, it is unfortunate that these param- eters were not measured. Generally, toxicity agreed with expectations based on the chemical composition of each effluent. The effluents differed both in the strength and the nature of their toxicities.

6. Implications for selector design and operation

With a successful selector operation, several operational issues have been identified and appropriate operator inter- ventions have been developed. The selector has proven effective in controlling filamentous bacteria levels and poly- saccharide levels have stayed within normal mixed liquor concentrations. Sludge bulking is well controlled and efflu- ent quality has been consistently good. Process adjustments were made based on regular wastewater characterization, microscopic examination of mixed liquor, (presence of filaments, globular zooglea and polysaccharides) and visual

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observations such as aeration tank foam and effluent quality. F/M and SRT together dictate the required selector size. Distributing the feed across the selector entrance was needed to lower F/M in the selector. Operating at a 2 < F/M < 5 on COD basis, on the other hand, requires proportionally higher reactor volume for a given SRT. Therefore, results indicating slow gradual changes because of long response times at SRT as high as 18 days, are achievable. Solids deposition prevention in sumps and the equalization tank just before the selector stage may improve settleability and reduce VFAs production without commensurate selector tankage requirements. However, minimization before the selector would not affect any potential reaction-limiting effects inside the selector. Tem- perature is an important design consideration for the selec- tor process which may be critical depending on the regional climate. Increased VFAs production at a high temperature during summer season may require additional tankage, higher SRT, and lower solids concentration to reduce VFA production. An important observation, was the tendency of solids to float in the equalization section of the selector system. Flo- tation appeared to be caused by gases produced within the sludge blanket. Floating solids would accumulate until they either plugged the submerged launder or escape into the effluent where they occasionally plugged the submersible pump that conveyed supernatant to the selector unit. A provision of an adequate float solids collection and removal-recycling system would be required. The rapid buildup of grease in influent sludge piping, was another noteworthy operational problem observed in the plant. During piping repairs, grease buildup that had reduced the effective sludge piping diameter was observed. Provi- sions to facilitates regular pipeline cleaning and effective dissolve air flotation systems, should be considered at slaughterhouses with heavy grease loadings. Four additional considerations became evident during the full-scale-experiments include, DO, organic loading, toxicity, and wastewater characteristics variability. DO should be maintained at 4–6 mg/L residual in the selector to adequately match COD utilization rate. A taper aeration system to provide DO residual gradient from 4 at head end to 2 mg/L at discharge was installed. Excess organic load- ings should be step fed around the selector to minimize polysaccharide and globular zooglea growth. Adjustments should be made gradually and in small increments. Toxic chemicals (solvents, chlorination, alum, polymer, etc.) can kill or inhibit the selector biomass. The additional volume of the aeration and chemicals eliminations from the selec- tor were needed to eliminate aquatic toxicity in the treated effluent. Good communication between manufacturing and wastewater personnel to manage toxic chemical releases and prevent upsets are needed. The selector designs should be conservative and flexible to accommodate wide variations in wastewater characteris- tics. The key factor that the full-scale demonstration units

focused on was the liquid–solid separation step because of the limited underflow concentrations achievable using gravity separation and the desire to reduce VFAs produc- tion. Equalization solids are undesirable because these sol- ids include degradable solids that increase the loading on the selector system and inert solids that accumulate in the biological system and displace active biomass, thereby reducing capacity.

7. Conclusions

The combination of aerobic selector, step feed, increased aeration tank capacity, and tapered aeration has proven to be effective in controlling filamentous bacteria at Hawally slaughterhouse wastewater treatment plant. It was an inter- esting approach to overcome problems with floating sus- pended matter discharged from equalization tank, which could be eliminated without using chemicals. Furthermore, proper design of the selector includes providing the correct retention time under aeration, with the flexibility to change this as wastewater characteristics change. In this regard, a high aeration capacity is required so that DO concentra- tion is high enough to penetrate the floc structure and match COD utilization rate. In addition, minimum DO residuals of 2 mg/L are recommended for both, and 4 mg/L preferred when VFAs are present. RAS reaeration is a valuable tool to ensure complete digestion of stored food in the mixed liquor solids. A multi-cell selector can provide this function while also pro- viding operating flexibility and efficient maintenance capa- bility. Step feed should be used to maintain a low filament population to serve as a backbone for the bacteria floc and prevent effluent turbidity. Likewise, step feed maybe used to shunt excess COD to the aeration tanks to minimize the formation of globular zooglea and excess polysaccha- rides. It is also useful in controlling excessive nitrification in the aeration tanks. Tapered aeration permits setting up appropriate DO residual gradients along the length of aer- ation tanks. The process must be properly monitored and controlled. Otherwise, the process can eventually become unstable and effluent quality deteriorates. Finally, the Microtox acute toxicity test has been suc- cessfully used to measure the toxicity of wastewater in slaughterhouse wastewater effluent samples. Results indi- cated that initially the wastewater caused inhibitory effects to the system because the toxicity of many chemicals is more severe under low oxygen tensions. While the selector configuration helped eliminate effluent aquatic toxicity. The toxicity of the three effluents varied in strength and character and was generally explicable based on chemical data. However, toxicity tests revealed strong chronic toxic- ity in the conventional activated sludge system effluent. Future chemical analyses should include some measure of ionic strength and ion balance. Electrical conductance is the minimum requirement, however total dissolved solids, hardness, alkalinity and major ions should ideally be included. Future measurement of color should be added


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to the analyses because toxicity testing can be confused or complicated by highly colored effluents.


This research was funded by Grant No. EV07/00 from the research administration of Kuwait University.


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