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Chapter Two

LITERATURE REVIEW

2.1

Characteristics of Municipal Wastewater

The wastewater generated by a community is called sewage, which is a mixture of domestic wastewater, industrial wastewater (where the industry is discharging its wastewater in the same sewage system) and rain water, (where a single sewer systems exists for the wastewater and storm water (van Haandel and Lettinga, 1994). In the developing/underdeveloped countries of the world, more than 90% of the sewage is discharged untreated in the environment due to lack of proper wastewater collection and treatment facilities (Elliot, 1999; Niemczynowics, 1997). The quantity and strength of wastewater is governed by the size and socioeconomic status of the population of the community (Haskoning and Wageningen, 1994). Table 2.1 compares the characteristics of municipal sewage of Pakistani cities with sewage of various cities of the world. The composition of sewage varies greatly and its characterization is important for determining the size and designing of treatment plant (Haskoning and Wageningen, 1994; Henze and Ledin, 2001; Mahmoud, 2002). 2.2 Sewage Treatment

Anaerobic treatment is preferred to treat municipal wastewater because of its merits over conventional treatment methods (van Lier and Lettinga, 1999). These advantages are (i) its ability to treat high COD loads and withstand fluctuation in the influent, (ii) biogas formation, (iii) effective treatment of wastewater in a short period of time (James and Kamaraj, 2002). Anaerobic reactors reduce pollution load and provide good stabilization of solids. Furthermore, depending on the design of a UASB reactor, a high sludge hold-up time can be obtained so that the excess sludge be discharged only once every three to four years (Lettinga, 1996).

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Table 2.1 Composition of sewage in different cities of the world Parameters BOD COD Chlorides Sulphates NH4+-N Nkj-N Total P PO4-3-P TDS TSS VSS Temperature
1

Pakistan Karachi1 220-475 200-1400 300-1200 50-200 1000-1800 250-900 Lahore1 200-215 580-803 32-72 486-598 106-176 -

Palestine Al Bireh2 1586 80 104 13 12.9 736 617 -

Brazil Pedregal3 368 727 110 18 34 44 11 8 492 252 24-26

Columbia Cali3 95 267 17 24 1.3 215 107 24-27

Netherlands Bennekom3 231 520 15 18 14 8-20

All parameters are in mg/L except temperature (oC) Planning and Development Division of Pakistan (1987); 2Mehmoud (2002); 3 van Haandel and Lettinga (1994).

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2.2.1

Upflow anaerobic sludge blanket reactor process

The major achievement in the development of anaerobic treatment was the introduction of high-rate reactors in which biomass retention and liquid retention are not interlinked (Lettinga et al., 2001). Among the various anaerobic wastewater treatment technologies, upflow anaerobic sludge blanket (UASB) reactors have achieved considerable success and these reactors have been applied to treat a wide range of effluents such as sugar, pulp and paper, dairy, chemical, potato starch, bean balancing, soft drinks, fish processing, noodle processing, yeast production, slaughterhouse and coffee processing industries (van Lier and Bonez, 2001; Lettinga et al., 1997; Metcalf and Eddy, 2003). Wastewater to be treated is introduced from the bottom of the reactor and it flows upward through a blanket of biologically activated sludge, which is generally in the form of granular aggregates. The sludge aggregates have very good stability and do not get washed out under practical conditions. Therefore, provide good treatment efficiency when the wastewater comes in contact with the granules (Hulshoff Pol et. al., 1983; Hulshoff Pol, 1989). The gases (methane and carbon dioxide) produced under anaerobic conditions cause internal mixing, which helps in the formation and maintenance of biological granules. However, some of the gas produced in the sludge blanket is attached to the granules, and a gas- liquid-solid separator (GLSS) is added on the top of the reactor for the effective segregation of gas, liquid and granules. In GLSS, the gas surrounded particles strike with the bottom of degassing baffles and fall back into the sludge blanket and the treated water flows out of the reactor. The prominent physical features of a UASB reactor requiring careful consideration are the feed inlet, gas separation, gas collection, and effluent withdrawal. The feed inlet and gas separation designs are exclusive for the UASB reactor. The feed inlet is designed to provide a uniform distribution of wastewater and to avoid channeling or the formation of dead zones. There is lower gas production in sewage as compared to high strength wastewater, which leads to less circulation of gas to support the formation of biological granules. To control channeling is even more important for weaker wastewaters like sewage. Hence, a number of feed inlet pipes are used to direct flow to different areas of the bottom of the reactor from a common feed source (Metcalf and Eddy, 2003).

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2.3

Parameters Affecting the Efficiency of a UASB Reactor

The efficiency of a UASB reactor is governed by several process conditions, including temperature, pH, hydraulic retention time (HRT), organic loading rate (OLR), upflow velocity, type of seed sludge and sludge age (Wiegent, 2001; Vieira and Garcia, 1992). 2.3.1 Effect of temperature

The temperature considerably influences the growth and survival of microorganisms and three ranges of temperature are well recognized in biological treatment of effluents (Madigan et al., 1997). These ranges are termed as psychrophilic, mesophilic and thermophilic. Optimum temperatures for psychrophilic, mesophilic and thermophilic microorganisms are around 17oC, 35-40oC, and 55-60oC, respectively (Edeline, 1997). Although anaerobic treatment is possible at these temperature ranges, low temperature usually leads to a decline in the maximum specific growth rate and methanogenic activity (Bodik et al., 2000). Methanogenic activity begins in the minimum temperature range of 10-15oC. However, activity at this temperature range will be 10 to 20 times lower than the activity at 35oC, which requires an increase of the biomass in the reactor (10 to 20 times) or to operate at higher sludge retention time (SRT) and hydraulic retention time (HRT) in order to achieve the same COD removal efficiency as that obtained at 35oC (Kalogo and Verstraete, 2001; Foresti, 2001; Mahmoud, 2002). It is argued that the reduction in operational temperature not only retards the hydrolysis step but also leads to a significant decrease in the maximum growth and substrate utilization rates (Lettinga et al., 2001). A 33% drop in the soluble COD removal efficiency (i.e. from 53% at 20oC to 20% at 10oC) was reported by Van der Last and Lettinga (1992) when temperature was reduced below 10oC during anaerobic treatment of domestic sewage using an expanded granular sludge bed (EGSB) reactor. A decline in the efficiency of the EGSB reactor may owe to a reduction in methanogenic activity of the sludge or a limited acidification of the soluble COD fraction of sewage. Singh et al. (1996) treated municipal wastewater using a UASB system under low-temperature conditions and reported 70% COD removal at 11C and 30 to 50% at 6oC. Lew et al. (2003) found a gradual decrease in COD removal efficiency as the temperature was decreased. They reported 82% COD removal at 28C, 72% at 20C, 68% at 14C and 38% at 10C. Kalogo and Verstraete

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(2001) also found that COD removal efficiency at temperature in the range of 10-15oC was lower than that of efficiency at 35oC. Van Lier and Lettinga (1999) studied the effect of transient temperature rise on the performance of a UASB reactor containing mesophilic microorganisms. There was an increase in the methane production with an increase in the temperature due to the accelerated methanogenic activity. However, a sharp drop in the methane generation was noted at the reactor temperature exceeding 45oC because of a substantial decline in the activity of mesophilic granular sludge due to bacterial inactivation. Halalsheh (2002) treated high strength sewage (COD = 1531 mg/L) using a UASB pilot plant under subtropical conditions. The CODtot removal efficiencies were 62% and 51% in summer and winter, respectively when the plant was operated at ambient temperature (18-25oC) and hydraulic retention time of 24 hours. 2.3.2 Effect of pH

The pH of an anaerobic reactor is extremely important because methanogenesis process can proceed at a high rate only when the pH is maintained in the range of 6.3-7.8. In the case of domestic sewage, pH naturally remains in this range because of the buffering capacity of the acid-base system (carbonate system), and addition of chemical is not required (van Haandel and Lettinga, 1994). The UASB reactors employed for sewage treatment in tropical and subtropical countries are reported to be extremely stable in terms of pH and buffering capacity (Leitao, 2004; Seghezzo, 2004; Mgana, 2003; Cavalcanti, 2003; Halalsheh, 2002). Zhang et al., (2005) found improvement in both hydrolysis and acidogenesis rates while treating kitchen wastewater using a two phase anaerobic reactor and reported that pH 7 provided an optimal working environment for anaerobic digestion resulting into TOC and COD removal of 86% and 82%, respectively. 2.3.3 Effect of hydraulic retention time (HRT) and upflow velocity (Vup)

The HRT is one of the most important parameters affecting the performance of a UASB reactor when used for the treatment of municipal wastewater (Vieira and Garcia, 1992). The upflow velocity is directly related with HRT and plays an important role to entrap

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suspended solids. A decrease in Vup entails an increase in HRT, which boosts SS removal efficiency of the system (van Haandel and Lettinga, 1994). The COD removal efficiency of a UASB reactor also decreases at elevated upflow velocity because higher Vup reduces the contact time between sludge and wastewater in addition to smashing of sludge granules, and resultantly higher washout of solids (van Haandel and Lettinga, 1994; Gonclaves et al., 1994; Kalogo and Verstraete, 1999; Mehmoud, 2002; Leitao, 2004). However, Vieira and Garcia (1992) reported no distinct effect of HRT on the treatment efficiency when a reactor was operated at hydraulic time in the range of 4.4-14.5 hours with Vup varying between 0.4 and 1.3 m/hr. According to Halalsheh (2002), COD removal efficiency of the reactor was only 62 % at a longer HRT of 24 hrs and at temperature of 25oC. The difference of opinion in scientific community is may be due to the difference in reactor design, operating procedures and range of HRT. UASB treatment plants are reportedly working in various tropical cities, such as Sao Paulo and Saumare in Brazil, Kampur and Mirzapur in India and Bucaramanga and Cali in Columbia (Vieira et. al., 1994; Draaijer et al., 1992; Tare et al., 1997; Schellinkhout and Collazos, 1992; Schellinkhout and Oscario, 1994). The HRT in these plants varied between 4-20 hours but was mostly in the range of 4-8 hours (Cavalacanti et. al., 1999). The results demonstrated that COD removal efficiency of the reactor was a function of HRT and COD removal efficiency approached to 80% even though the sludge was not well granulated and was merely a suspended fluffy mass (Kalogo and Verstraete, 1999). However, too short HRT causes a rapid decline in the performance of UASB reactor (Ragen et al., 2001). For instance, Leitao (2004) reported 93% SS and 60% CODtot removal efficiency of a UASB reactor at HRT of 6 hrs. Below this HRT, efficiency of the reactor dropped significantly, and SS removal efficiency of the reactor was almost negligible at HRT of 1 hr. Fang (2000) studied the effect of HRT on the acidogenesis of dairy wastewater at 37oC temperature and retention time in the range of 4 to 24 hrs. He found an increase in acidification from 28 to 54% by increasing HRT from 4 to 12 hrs, beyond which there was no increase in the acidification. Tronovec and Britz (1998) treated food industry wastewater by using a UASB rector and reported COD removal efficiency of the reactor more than 90% at HRT of 10 hrs.

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2.3.4

Effect of organic loading rate (OLR)

The OLR can be changed by varying the COD concentration of influent, flow rate, HRT and volume of the reactor in accordance with the following relationship.

OLR

Q COD COD== + V

COD HRT

Where OLR = Organic loading rate (kg COD/m3.day) COD = Total chemical oxygen demand of influent(kg COD/m3) Q = flow rate (m3/day) V = reactor volume (m3) HRT = Hydraulic retention time (days) The effect of OLR on the performance of a UASB reactor is yet not clear as it depends on a number of factors (shown in above relationship), which have a dissimilar effect, mostly contradictory, on the performance of UASB reactor (Leitao, 2004). Some researchers reported an increase in the efficiency of high rate anaerobic reactors with increasing OLR (Ruiz et al., 1997; Kalyuzhnyi et. al., 1998; Brown, 1998). However, that increase was upto a certain OLR, beyond which there was sludge bed flotation and excessive foaming in the gas-liquid-solids separator (GLSS). Higher OLR also results in the accumulation of biogas in the sludge bed forming gas pockets that ultimately cause sludge flotation. Leitao (2004) determined the effect of organic loading rate OLR on the COD removal efficiency of three UASB reactors employed to treat sewage. The reactors were operated for 6 hrs to treat effluents varying in COD contents and OLR. In the case of effluent having COD contents lower than 300 mg/L, the efficiency of UASB reactors was low. However, reactors showed maximum COD removal efficiency of 60% when the COD concentration was higher than 300 mg/L. He further reported a decrease in SS removal efficiency of UASB reactors with an increase in OLR (Leitao, 2004). When the increase in OLR is due to an increase in the influent COD contents, a sharp decrease in SS removal efficiency may occur. That

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decrease owes to SS washout caused by turbulence due to higher rate of gas production. When the increase in OLR is associated with decreasing HRT (increase of flow rate), a decline in SS removal efficiency may occur due to sludge washout and short contact time between sludge bed and substrate, which results into the malfunctioning of physical and biological processes taking place in the reactor. However, SS removal efficiency of the reactor slightly decreases when increase in OLR is associated with increased SS contents of substrate (Leitao, 2004). In contrast, Miron (1997) reported an increase in SS removal with an increase in OLR associated with higher influent SS. These differences among the research workers may owe to use of primary sludge by Miron to increase OLR. Primary sludge is mainly comprised of settleable suspended solids which increased the SS removal efficiency (Miron 1997, Leitao, 2004). 2.4 Post Treatment of a UASB Reactor Treated Wastewater

Anaerobic reactors have an inherent disadvantage that microbial quality of the effluent does not comply with the irrigation standards established by the environmental agencies (Chernicharo et. al., 2001; von Sperling et al. 2002, 2004; Chernicharo, 2006). Therefore, post treatment is required for the removal of pathogens from anaerobically treated effluent, considering the public health risk and use of treated water for irrigation. Factors affecting the disinfection efficiency of a system include the type of disinfectant, type and number of microorganisms in wastewater, quality of wastewater, pH and temperature (Kuo et al., 1997; Lazarova et al., 1998; Collivignarelli et al., 2000). Post treatment is accomplished by using both conventional and advanced oxidation processes. Conventional treatment systems include maturation ponds, solar UV irradiation, chlorination (USEPA, 1986) polishing ponds (von sperling and Mascarenhas, 2004), waste stabilization ponds (van Haandel and Lettinga, 1994), constructed wetlands (Sousa et al., 2001), overland flow system (Chernicharo et al., 2001) rotating biological contactors (Tawfik et al. 2002, 2003), downflow hanging sponge (Tandukar et al, 2005 Machdar et. al., 2000, Uemura et. al., 2002). In contrast, advanced oxidative processes (AOPs) can be potentially used for the inactivation of a wide spectrum of pathogenic microorganisms (Machado et. al., 2007; Assalin et. al, 2004). AOPs are preferred over conventional treatment techniques because of several distinct advantages (Zhou and Smith, 2002; Hunt and Marinas, 1997). AOPs are

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based on the production of secondary radicals (OH-) which are considered as the most reactive oxidizing agents in water treatment that can be used both for the oxidation of organic and inorganic compounds and disinfection of microorganisms (Glaze, 1987; Gulyas et al., 1995; Beltran et al., 1999). AOPs include O3, H2O2 alone and in

combination. Post treatment technology however needs to be reliable, efficient, plain in construction, operation and maintenance, able to withstand fluctuations in hydraulic and organic shock loads and energy efficient (Tai et al., 2004). 2.4.1 Disinfection with sunlight

A number of researchers have recognized sunlight radiation capable to damage microorganisms (Acra et al., 1990; Joyce et al., 1992; Reed, 1997) and to effectively inactivate sewage microorganisms like Enterococci or Escherichia coli (Rozen and Belkin, 2001; Sinton, 2002). Sunlight produces highly reactive forms of oxygen species in water. These reactive molecules contribute in the destruction process of the microorganisms. Solar disinfection can be effective against pathogens in UASB reactor treated effluent, especially in countries where eight hours of continuous sunlight is available throughout the year. The components of solar spectrum responsible for the inactivation of different microorganisms include UV-B (290-320 nm), UV-A (320-400 nm) and blue to green visible light (400-550 nm). Sunlight disinfection of microorganisms can be explained in terms of three main mechanisms; (i) direct absorption of solar UV-B energy, which damages DNA, this process is oxygen- independent, (ii) Photo-oxidation catalyzed by endogenous photo-sensitizers resulting in the generation of reactive oxygen species that damage internal cell targets like strands of DNA and (iii) Photo-oxidation catalyzed by exogenous photo-sensitizers which damages external structures such as cell membranes (Webb and Brown, 1979; Whitelam and Codd, 1986). According to Davies-Colley et al. (1997), all these components of sunlight contribute equally in the inactivation of Enterococci and F- RNA phage. Whereas, UV-B energy irradiation dominates the inactivation of E.coli and F-DNA at moderate pH. However, the repair mechanism in microorganisms may be able to cope with the damage inflicted due to lower dose of UV irradiation (Calkins et al., 1976). Disinfection by sunlight is influenced by factors including oxygen content of water, pH, temperature and turbidity (Davies-Colley et al., 1997; Reed, 1997). Davies-Colley et al. (1999) reported that sunlight (seven hours

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exposure) dominated the inactivation of fecal colifrom in waste stabilization ponds treated effluents. 2.4.2 Disinfection with hydrogen peroxide

Hydrogen peroxide is very effective for the removal of microorganisms, and provides long term disinfection. Its advantages over other oxidants are its non toxic, harmless and environment friendly by-products (Tofant et al., 2006). The proficiency of hydrogen peroxide to disinfect wastewater is due to the production of powerful oxidants, such as nascent oxygen, super oxide radicals and the hydroxyl radicals that are highly reactive and toxic for microorganisms (Imlay et al., 1988; Davidson and Branen, 1993; Ksibi, 2006). According to Imlay et al. (1988), these radicals have both lethal and sub lethal effect on bacterial genes and cause permanent damage to host cell components and DNA causing physiological alteration, growth delay and oxidative disturbances of bacterial membrane. There is reportedly a complete membrane collapse after 17 minutes of exposure to hydroxyl radicals (Anzai et al., 1999; Davidson and Branen, 1993). Toledo et al., (1973) determined the effect of temperature on the antimicrobial efficacy of H2O2. They observed a notable increase in the sporicidal activity of H2O2 with increased temperature. Ksibi (2006) reported high level of bacterial inactivation with a H2O2 dose of 2.5 ml/L. Yasar et al., (2007) found a H2O2 dose of 170 mg/L to be optimal to disinfect the UASB treated industrial effluent as it provided more than 99% removal of the pathogens like salmonella, E-coli, fecal Coliform and fecal Streptococcus. 2.4.3 Disinfection with ultraviolet irradiation

UV irradiation has been widely used for disinfection since long due to its advantages such as its non toxic effect due to no production of by-products, simplicity, minimal space requirement, no use of chemicals, short contact time and lower cost than chemical treatment (USEPA, 1999; Collivignarelli et. al., 2000; Blatchley et. al., 2001; Tessele et al., 2005; Petri et al., 2006). The process involves the use of low pressure UV lamps with a principal wavelength of 254 nm. The wavelength that shows maximum disinfection rate is in the range of 250-270 nm (Bukhari et al. 1999; Darby et al. 1994). Unlike ozone, UV irradiation inactivates the bacterial cell by penetrating the cell wall and altering the nucleic acids (DNA

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and RNA) of microorganisms. However, higher UV dose may lead to the disruption of cell membrane and eventually death of cell (Wolfe, 1990; Braunstein et al., 1996; Oppenheimer et al., 1997; Lazarova et al., 1998). Caretti and Lubello (2003) find a UV dose of 330 mJ/cm2 effective for the inactivation of both fecal coliform and E.coli. However, disinfection by UV is a function of various parameters like UV transmittance, exposure time, quality of treated wastewater and reactor configuration (Zhou and Smith, 2002). The suspended solids and pollutants like fats, oil and colored compounds substantially reduce the intensity of radiation received by microorganisms (Whitby and Palmateer, 1993). White (1986) recommended suspended solids (SS) concentration of 20 mg/L or lower of wastewater for the application of UV. The content of suspended solids, especially of large particles, has a strong impact on UV disinfection efficiency. Large particles (greater than 10 m diameter) absorb the UV light and also shield the microorganisms from UV exposure. Therefore, large particles associated fecal coliforms are difficult to be disinfected (Wang et al., 2006). It is considered a disadvantage of UV irradiation that some microorganisms repair damaged DNA in a few days after exposure to UV irradiation because UV treatment does not provide residual protection. A higher UV dose however, hampers the repair mecchsnisn of microorganisms (Lindenauer and Darby, 1994, Tosa and Hirata, 1999). 2.4.4 Disinfection with ozone Ozone is now considered an effective bactericide, paracitiside and virucide (Summerfelt and Hochheimer, 1997; Liltved and Landfald, 1995). Ozone is a strong and highly reactive oxidant due to its high redox potential, and is used to disinfect wastewater (Tomiyashi et al., 1985; Rice, 1996). It kills microbes by oxidizing the lipid bi-layer and damaging the nucleic acid compounds such as purine and pyrimidine. The disinfection performance of ozone is a function of ozone dose, contact time and quality of wastewater to maintain a residual ozone concentration (Langlais et. al., 1991; Sharrer and Summerfelt, 2007). Ozone attacks microorganisms in two ways (WEF, 1996). In acidic environment, molecular ozone reactions are slow, selective and microorganisms undergo direct ozone attack. At elevated pH, the efficiency of ozone is increased due to rapid production of OH- radicals, which are highly reactive and non selective (Stachelin and Hoigne, 1985; Rakness et al. 1993; Gehr et al., 2003). However, inactivation of microorganisms is

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generally attained by the action of molecular ozone than free radicals (Youteri and Gurol, 1988; Zhou and Smith, 2002). The dissolved substances and suspended solids considerably reduce the disinfection capacity of ozone (Stachelin and Hoigne, 1985; Zhou and Smith, 2002). The advantage of ozone application is that it produces less undesirable reaction by-products (Yuteri and Gurol, 1987, 1988; Zhu et al. 1989). An additional benefit is that dissolved oxygen concentration of treated effluent is elevated to near saturation levels as ozone rapidly decomposes (Metcalf and Eddy, 1991). Although ozone provides high level of disinfection, regrowth and recovery of microorganisms can take place in treated water (Lindenaver and Darby 1994; Braunstein et al. 1996). 2.4.5 Disinfection with combination of AOPs

Combined application of AOPS may enhance the microbial inactivation due to synergistic oxidation effect (Zhou and Smith, 2002), and the efficiency of combined disinfection method is greater than the efficiency of disinfectants (Lubello et al., 2002; Caretti and Lubello, 2003). The integrated application of different AOPs causes multiple damage to microorganisms. In case of a single disinfectant, the damage may be smaller and susceptible to repair, which may not be possible in case of two disinfectants causing a greater variety of damages. The combinations of two disinfection methods can also destroy a wider rage of microorganisms as compared to a single disinfection technique. It may be more efficient and economical to apply a combination of low chemical and UV doses, instead of a high dose of one disinfectant. 2.4.5.1 Application of O3/H2O2 process (peroxone)

The O3/H2O2 process, also called peroxone, has been commonly used to disinfect wastewater due to its simplicity and cost effectiveness. Peroxone has been reported to be used as an effective germicide and is mainly used for the oxidation of micro pollutants, removal of pesticides and control of taste and odor causing compounds (USEPA, 1999; Paillard et al. 1989; Karimi et al. 1997). The combination of ozone and hydrogen peroxide is advantageous because H2O2 enhances the ozone decomposition and subsequently increases the yield of hydroxyl radicals in addition to increasing ozone mass transfer to liquid resulting in an improved ozone reaction rate (Dugnet et al., 1990; Glaze, 1987).

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2.4.5.2

Application of O3/UV process

The O3/UV process makes use of UV photons to activate ozone molecules, thereby promoting the formation of hydroxyl radicals (Peyton and Glaze, 1982, 1988), which are very effective in the reduction of microbial populations (Glaze, 1987; Venosa et al., 1984). The UV photolysis of ozone also yields H2O2 which can be additionally photolyzed to form two OH- radicals. The O3/UV process is difficult to use on a large scale but may be useful for small water and wastewater treatment plants, especially when the substrate has high UV absorbance (Glaze et al., 1987). It has been accepted that ozone can increase the UV transmittance of water (Benjamin et al., 2006; Mackey et al., 2004). According to Parker and Darby (1995), this process is effective in disinfecting the microorganisms as ozone lowers the turbidity and color of wastewater thereby enhancing the efficiency of UV irradiation. The O3/UV process is considered less economical as compared to O3/H2O2 process because the generation of H2O2 by the UV photolysis of ozone is much less efficient than the electrochemical method applied in industry (Peyton and Galze, 1988; Zhou and Smith, 2002). 2.4.5.3 Application of H2O2/UV process

The application of UV in combination with H2O2 accelerates the oxidative degradation due to the production of highly reactive hydroxyl radicals (Baxendale and Wilson, 1957). When UV light is absorbed directly by H2O2, two hydroxyl radicals are generated by the photolysis of peroxidic bond. These hydroxyl radicals further undergo a decomposition and formation cycle (Crittenden et al., 1999). These hydroxyl radicals are generated efficiently when UV irradiation is in the wavelength range of 200-280 nm (Baxendale and Wilson, 1957). Determination of an optimal H2O2 dose is vital for this process because an adequate dose of H2O2 is required to absorb the UV and generate hydroxyl radicals. Higher dose will hamper hydroxyl radical generation due to the scavenging nature of H2O2 (Zhou and Smith, 2002). Ahn et al., (2005) reported that with an increase in the dose of H2O2, shorter UV contact time was required for the removal of total coliform and a dose of 50 mg/L of hydrogen peroxide with a UV contact time of six minutes was effective to meet the criteria for total coliform removal.

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2.5

Reactivation of Pathogens in Disinfected Wastewater

Regrowth potential of pathogens needs to be studied for a sustainable use of disinfected wastewater for irrigation. According to Sommer et al., (2000), microbial DNA is capable of photreactivation after exposure to low pressure UV irradiation as this process does not provide a disinfectant residual. There are two mechanisms for the repair of DNA i.e. (i) nucleotide excision repair usually referred to as dark repair and (ii) photoreactivation. In case of dark repair mechanism, a strain of DNA synthesizes a copy of DNA by the action proteins in order to replace the damaged DNA strain. In the photoreactivation, a single enzyme called photolyase reverses the damage caused to DNA by UV irradiation. Photo reactivation process depends on the presence of light in the wavelength range of 300 to 500 nm to complete the repair process (Friedberg et al., 1995; Thoma, 1999; Zhou and Smith, 2002). There is an increase in the regrowth of microorganisms in ozone disinfected effluent due to the production of biodegradable by- products such as aldehydes and ketones which act as nutrients (Langlais et al., 1991). The rate of regrowth is a function of (i) types and amounts of nutrients present in treated effluent, (ii) residual dose of disinfectant and (iii) corrosion products formed during storage (Lindenauer and Darby, 1994, Escobar and Randall, 2001; Alonso et al., 2004). The repair mechanism of microorganisms can be hampered by applying higher UV dose (Tosa and Hirata, 1999). Alonso et al., (2004) performed a series of experiments using UV irradiation and ozone for the removal of fecal coliforms, fecal streptococci, salmonella spp, and clostridium from wastewater of a treatment plant. They reported 99% removal of fecal coliform, salmonella and fecal streptococcus using Ozone and UV irradiation. Regrowth was more prominent in UV disinfected wastewater as compared to ozone. The efficiency of ozone was better because ozone causes destruction of cell whereas UV irradiation hampers the ability of microorganism to replicate. According to Yasar et al., (2007), temperature is one of the most important factors in the regrowth phenomenon in addition to the dose, residual concentration of disinfectants and genetic characteristics of species. 2.6 Restart of UASB Reactor after Shutdown Shutdown of UASB reactors is common in the case of small municipal treatment plants and seasonally operated industries. The restart of a UASB reactor is however a complex process because of the sensitivity of methanogenic microorganisms in the sludge bed to changes in

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the temperature, organic loading rate and other external factors that can cause delay in the response of microbial population (Dong et. al, 2010). Therefore, the restart performance of a UASB reactor requires determining its ability to endure long shutdown period. It has been reported that the performance of UASB reactor after a short non feeding period (upto four months) is not affected considerably in terms of pollutant removal and methane production rate (Young, 1980; Manariotis and Grigoropoulos, 2006; Dong et. al., 2010). Young (1980) reported that the BOD removal efficiency of a USB reactor was 50% immediately after restart and increased to 90% and 95 % respectively after 16 and 24 days of restart following a six months shutdown. Similarly, Sanz and Fdz-Polanco (1989) reported that the normal functioning of the UASB reactor treating municipal wastewater was restored after 25 days of restart with a COD removal efficiency of more than 72 % after six months shutdown period. However, after a long non feeding period the performance of UASB reactor can be affected substantially and varies considerably as it is dependent on the history of the reactors, type of packing material/sludge used and type of wastewater treated (Manariotis and Grigoropoulos, 2008). According to Manariotis and Grigoropoulos (2008), a period of 2.5 months was required to restore the performance of anaerobic reactors after a long shutdown period of 24 months. Elevated temperature is found to affect the methanogenic activity of microbial population in the UASB reactor and leads to an increase in the time period required to restore the normal functioning of the reactor after restart. Lepisto and Rintala (1995) reported a decrease in the specific methane production rate and increase in time period required for restoring normal performance of a UASB reactor at 70oC following a shutdown period of two weeks. However, after a shutdown period of 6 days, there was no affect on methane production rate.