A Brief Report on

Wastewater Treatment and Management Project

in Asian Regional Research Project in Environmental Technology (ARRPET) Phase II

Funded by Swedish International Development Cooperation Agency (SIDA)

Co-coordinated by Asian Institute of Technology

Prepared by: Saumyen Guha

Department of Civil Engineering Indian Institute of Technology Kanpur, UP 208016, INDIA

with inputs from the NRIs of WWTM Project

August, 2007

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Table of Contents
Outline of the Report __________________________________________________3 Background _________________________________________________________3 Research ____________________________________________________________4 Networking __________________________________________________________5 Dissemination________________________________________________________6 Capacity Building_____________________________________________________6 Administration _______________________________________________________6 AIT Report: Efficient Processes for Wastewater Nitrogen Removal _____________7 CENTEMA Report: Sustainable Development of Tapioca Processing Industry in Vietnam____________________________________________________________15 IITK Report: Domestic Wastewater Treatment in India: Optimization of UASB Reactor ____________________________________________________________25 IITB Report: Characterization of Biosolids fromUASB/Activated Sludge Process/Fluidized Bed Process _________________________________________32 KMUTT Report: Bioremediation and Reuse of Marine Shrimp Farm Effluent __40

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Outline of the Report

First four pages of this report provide an executive summary of the WWTM project in ARRPET Phase II. This is followed by a brief report from each NRI on the progress and achievement of the individual projects during Phase II.

Background
The main goal of the Wastewater Treatment and Management Project (WWTM) was to develop cost effective technologies for the region using local resources and expertise for the treatment of wastewater originating from cities, towns and villages of the region. Researchers (NRIs) from several countries work on different project segments in order to meet the overall goal of the project. At the inception of the project in Phase I, there were 7 NRIs from 4 different countries namely India, Thailand, Vietnam and Malaysian. During the transition from Phase I to Phase II, one of the NRI from India and the NRI from Malaysia were discontinued on the basis of non-performance. Five NRIs in Phase II are: Asian Institute of Technology (AIT), Thailand. e-mail: ajit@ait.ac.th Centre for Environmental Technology and Management (CETEMA), Van Lang University, Vietnam. e-mail: centema1@hcm.vnn.vn Indian Institute of Technology Kanpur (IITK), India. e-mail: sguha@iitk.ac.in Indian Institute of Technology Bombay (IITB), India. e-mail: cshnath@iitb.ac.in King Mongkuts University of Technology (KMUTT), Thailand. e-mail: piyabutr.wan@kmutt.ac.th

In terms of pollutants, the project in Phase II targets carbon and nitrogen removal. The distribution of NRIs according to the pollutants in Phase II is: Carbon Removal: IITK, IITB and CENTEMA Nitrogen Removal: AIT and KMUTT

Based on the local economy, the wastewater may consist of pure domestic sewage, domestic sewage mixed with small-scale industry wastewater and agro-industry wastewater. The distribution of NRIs according to the wastewater is: Domestic Wastewater: IITK and IITB Agro Industry Wastewater: CENTEMA Shrimp and Seafood Industry wastewater: AIT and KMUTT

NRIs are working on a diverse range of unit processes such as Upflow Anaerobic Sludge Blanket Reactor system for domestic and agro industry wastewater (IITK, IITB and CENTEMA), photo-bio reactor system for shrimp pond wastewater (KMUTT) and nitrogen removal by anaerobic process (AIT). The major research achievements are summarized in the next section.

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Research
The research in ARRPET consisted of a combination of development of novel technology and adaptation of the existing unit processes to local environmental problems. While developing the technologies, special attention was given to utilization of low cost and locally available materials and expertise. The research started in Phase I of ARRPET. In Phase II, all the NRIs were conducting laboratory experiments with real wastewater and successfully demonstrated the applications in Pilot Scale treatment plants. Funding for the pilot plant construction, operation and maintenance were secured from private partners in most cases. Cost Benefit Analysis was conducted by each of the NRIs for their respective projects. All the projects were proved superior compared to the benchmark technologies. Benchmarks were selected based on frequently used technologies used in the area for the target wastewater. Brief outlines of the research achievements of each NRI are presented below: AIT: A novel process called combined Activated Sludge with Partial Nitrification (AS/PN) was coupled with Anaerobic Ammonium Oxidation (ANAMMOX) process to remove nitrogen from the wastewater. Target wastewater in this case was effluent after the secondary treatment of sewage and from seafood industry. Several experiments, both in laboratory and pilot plant scale, were conducted to develop the novel process, which can simultaneously and efficiently remove both organic carbon and nitrogen pollution. In the laboratory experiments, maximum nitrogen removal rate was 1.4 kg N m-3 d-1 at ammonium and nitrite concentrations of around 375 and 350 mg N l-1, respectively. The experiments demonstrated that combined AS/PN ANAMMOX process is stable for wastewater containing high ammonium up to 700 mg N/l. Pilot scale experiments with real wastewater from seafood industry also showed satisfactory results. CENTEMA: Goal of this project was to develop a model for the sustainable development of tapioca industry at the village level through waste minimization, reprocessing and recycling. Proposed wastewater treatment system consisted of the UASB reactor system combined with an aerobic activated sludge process and stabilization ponds. The treated wastewater was reused for irrigation in the area for cassava cultivation and on the surrounding agricultural land. The wasted sludge from wastewater treatment plant and cassava root peel was used for composting. The compost produced was applied for the cassava and corn cultivations. The fish produced in the stabilization ponds were edible and was a source of revenue. In this way, the target was to create a zero waste industrial ecosystem for the tapioca processing villages, where solid waste and wastewater can be gathered and treated at the central treatment units in the area. IITK: A field and literature survey during Phase I of ARRPET indicated that the UASB process for the treatment of low strength wastewater suffered from the following shortcomings: longer start-up times, inability to form self-immobilized bacterial granules and very low bio-gas recovery. Primary objective of the project at IITK was to achieve better granulation in UASB reactors treating low-strength wastewater. This was achieved using locally available and low cost natural polymer additives such as Reetha seed (Sapindus trifoliata) extract and Chitosan. It was also shown that the enhanced granules were stable for long period under various shock loading without requiring further doses of the additives. Large granules were

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generated in the reactor using real sewage with COD concentration varying randomly between 100-200 mg/L. More than 80% COD removal could be achieved with an HRT of only 2.5 hours in the sludge bed and 4.5 hours overall including the settling zone. The 100 m3/day pilot plant has been funded by Vapi Waste and Effluent Management Co. Ltd. (VWEMCL) and is operated jointly by IITK, IITB and VWEMCL. IITB: Enhanced granulation in UASB reactor with low strength wastewater, as described in IITK above was conducted as a collaborative project between IITK and IITB. While the reactor operation and monitoring of chemical and biochemical parameters were conducted at IITK, the granule examination and characterization using Image Analysis (IAS), Environmental Scanning Electron Microscope (ESEM), Electron Probe Micro Analysis (EPMA) and Transmission Electron Microscope (TEM) were conducted at IITB. Close collaboration between the two institutes enabled development of an applicable technology. The pilot plant for the enhanced granulation is also being operated jointly between IITK, IITB and VWEMCL. In addition, IITB acted as a hub for bio-solids characterization for all the NRIs in WWTM project and IHWTM project in ARRPET. KMUTT: This project attempted to develop cost-effective methods for removing ammonia nitrogen from shrimp pond water prior to discharge to the environment by utilizing photosynthesis activity of micro-algae developed in shrimp-pond. The key was to determine the light/dark period and mixing condition for optimum nutrient removal by green algae (Chlorella sp.). It was observed that the light/dark ratio and mixing are closely related parameters. In a shallow well-mixed tank, the light requirement is less compared to a deep tank with less mixing. In the laboratory scale experiments, maximum ammonia nitrogen removal rate observed was 0.71-0.98 mgN/mg-Chl-a/h. At the pilot scale the nominal ammonia nitrogen removal rate was 0.06 to 0.15 mg-N/mg-Chl-a/h which increased substantially to 0.50 mg-N/mg-Chla/h with mixing, bringing it close to the laboratory reactor observation. Treatment of the sediment from the shrimp pond was considered as the minor issue and preliminary investigations were conducted for biogas production, nutrient leaching and fate of microbial population during the drying process.

Networking
Networking was achieved at three different levels: (i) research networking through collaboration between two or more NRIs; (ii) transfer of technology and development of expertise through specialized workshops; and (iii) research collaboration between NRIs and industry partners through pilot scale experiments. NRIs from WWTM project as well as IHWTM project of ARRPET participated in the first two activities, which lead to healthy interaction and synergy between two projects in ARRPET. By close interaction between the NRIs, many research problems were resolved. Some of these are: characterization of biosolids, analytical methods for target compounds and metabolites, analysis of microbial ecology, identification and characterization of phytochelatins in phytoremediation, etc. The NRIs from two projects meet every year in a combined regional workshop to foster networking. Eight specialized training workshops have been organized so far for the NRIs in WWTM. Besides, there were 6 more training workshops for the industry partners. Collaboration with industry

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partners through pilot plant opened up the possibility for future transfer of technology developed in ARRPET.

Dissemination
Knowledge dissemination from ARRPET project was conducted at multiple levels. The technology developed was published in peer-reviewed journals and were presented at national and international conferences. Twelve articles in peer-reviewed journals have already been published and 8-10 more are in the process of preparation or review. A total of 14 publications have come out so far in the National and International conferences. This enabled technology evaluation by the peer groups at large as well as dissemination of the knowledge to technical community. Besides, 4 national workshops and were held by the NRIs in order to keep the stake holders and policy makers informed about the technologies developed at ARRPET. The NRIs from WWTM and IHWTM projects published two newsletters every year, which was circulated widely through internet. These articles were targeted towards the nontechnical community.

Capacity Building
Capacity building activity was mainly targeted towards building technical manpower. To this end, most of the NRIs had large success by guiding Masters and Ph.D. students as part of the project activity. Some laboratory infrastructure was built from the funding of Minor Equipment in the project.

Administration
Administrative structure of WWTM project consist of a Principal Investigator (PI) designate from amongst the NRIs whose duty is to monitor the NRI projects on a regular basis in consultation with a Swedish expert, give timely feedback to the NRIs, enforce the project requirements and prepare the project reports. The financial matter is handled by AIT. There was a change of PI in the WWTM project in the year 2004 and a change of Swedish expert in the year 2006. Brief reports from the NRIs of individual projects follow.

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AIT Report: Efficient Processes for Wastewater

Nitrogen Removal
Contact Person: Prof. Ajit P. Annachhatre Environmental Engineering and Management Asian Institute of Technology PO Box 4, Klong Luang Pathumthani 12120, THAILAND e-mail: ajit@ait.ac.th Technology Developed Through Research

Technology Developed in ARRPET II

ARRPET Phase I brought out that complete removal of nutrients such as nitrogen and phosphorus from domestic wastewater could not be achieved through UASB process. As a result, UASB effluent needs to be further treated for removal of nutrients. Accordingly, researches were undertaken in ARRPET Phase II for removal of nitrogen from the UASB effluent. The main objective during ARRPET II research was to develop an appropriate and efficient biological nitrogen removal processes following Upflow Anaerobic Sludge Blanket (UASB) process. Accordingly, the research was initiated with a focus on high strength nitrogen wastewater with low organic carbon source. Extensive literature review has been carried out (Khin and Annachhatre, 2004). Several novel biological nitrogen removal processes were identified including Single reactor system for High Ammonia Removal Over Nitrite (SHARON), Anaerobic Ammonium Oxidation (ANAMMOX) and Completely Autotrophic Nitrogen Removal Over Nitrite (CANON). However, these processes require wastewater with very low organic carbon i.e. COD and BOD less than 100 mg/l. In reality, effluent from the UASB process rarely achieves this limit. One of the classical methods is to apply conventional nitrificationdenitrification process. However, this requires complete conversion of ammonia to nitrate and from nitrate to nitrogen gas which results in high oxygen demand. Furthermore, addition of external carbon source is required to allow denitrification to proceed. An alternative was to use combined SHARON-ANAMMOX process to treat the nitrogen pollution. However, SHARON process was known to be an autotrophic process, hence, a novel process has to be developed. By careful manipulation of pH and dissolved oxygen level during aeration, a simple activated sludge process (ASP) could actually perform partial nitrification similarly to a SHARON process. Extensive literature review as well as laboratory scale experiments have proved this hypothesis (Sinha and Annachhatre, 2006a; Sinha and Annachhatre, 2006b). Hence, a novel process called combined Activated Sludge with Partial Nitrification (AS/PN) and ANAMMOX process was developed. Several experiments, both in laboratory and

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pilot plant scale, were conducted to develop novel process which can simultaneously and efficiently remove both organic carbon and nitrogen pollution.

UASB Settling tank

Anammox gas-lift AS/PN Discharge

Influent Air

Figure 1: Schematic diagram of the developed technology (AS/PN-ANAMMOX process)

In essence, the developed technology comprises combined AS/PN-ANAMMOX process (Figure 1). Complete organic carbon removal takes place in the first aeration tank as well as partial oxidation of ammonium (only 50-60%) to nitrite. The Anammox process combines and converts ammonium and nitrite to nitrogen gas with a small amount of nitrate production (Eq. 2). AS/PN: ANAMMOX: NH4 + 1.5O2 NO2 + 2H+ + 2H2O NH4 + NO2 N2 + 2H2O Eq. 1 Eq. 2

Brief Description of Laboratory Research

Experiments of AS/PN and ANAMMOX processes were carried out with synthetic wastewater (high ammonium with no organic carbon source) in laboratory scale reactors. Several operating conditions (i.e. pH, DO and Temp.) have been tested and optimized. Effluent from AS/PN with almost equal amount of ammonium and nitrite was produced with the following operating conditions: pH: 7.5-8.5 Temperature: 35o C (in theory 25 40 o C is acceptable) DO: 0.5 mg/l The effluent from this reactor was fed into the Anammox reactor. The AS/PNANAMMOX process was operated successfully at the laboratory scale experiment. The maximum nitrogen removal rate was 1.4 kg N m-3 d-1 at ammonium and nitrite concentration of around 375 and 350 mg N l-1. The experiment demonstrated that combined partial nitrification-Anammox is suitable for high ammonium wastewater up to 700 mg N/l. The details of experimental results were included in Annual cum Progress Report 2006.

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Publications
Khin, T. and Annachhatre, A.P. (2004). Novel Microbial Nitrogen Removal Processes, Biotechnology Advances, 22 (7): 519-532. Hanh, D. N., Rajbhandari, B. K. and Annachhatre, A. P. (2005). Bioremediation of sediments from intensive aquaculture shrimp farms by using slow oxygen release agent. Environmental Technology, 26, 581-589. Sinha, B. and Annachhatre, A. P. (2006a). Assessment of partial nitrification reactor performance through microbial population shift using quinone profile, FISH and SEM. Bioresource Technology. (In press) Sinha, B. and Annachhatre, A. P. (2006b). Partial nitrification operational parameters and microorganisms involved. Rev. Environ. Sci. Biotechnol. (In press) Pilot Scale Application

Methodology
Following the successful operation in laboratory scale experiments, pilot plant investigation was initiated. Several industries have been contacted, however, none was willing to finance and install the developed technology at their plant. Finally, it was decided to bring wastewater from a seafood processing industry and carry out pilot plant investigation at AIT. Experimental setup for the whole pilot plant unit is illustrated in the Figure 2. It comprises AS/PN and Anammox gas-lift reactors. Details of each unit are described as follows: Activated sludge with partial nitrification (AS/PN) reactor setup: The 32 L CSTR was equipped with DO control as well as a mixer. Biomass was separated from the liquid in a 130 L stainless steel settling tank which has a scraper at the bottom. Effluent from this reactor was sent to Anammox process. Start-up and operation of this process was conducted using the seafood processing industry wastewater. Seed sludge was taken from the industry aeration tank. Temperature was not controlled and varied in the range of 27-36 oC.

Industrial wastewate r

AS/PN reactor

Settler

Effluent Anammox reactor

Wasted sludge

Figure 2: Process flow diagram of the pilot plant investigation

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Anammox reactor setup: The gas-lift reactor has a working volume of 32 L with an attached 12 L settler located at the top of the reactor. This reactor will receive effluent from AS/PN effluent. Start-up of the Anammox process was performed using synthetic wastewater as described previously (Progress Report No. 2). Seeding was performed by combining active Anammox biomass from previous research (1 L) with the activated sludge (4 L) from the industry. Anaerobic condition was achieved by flushing with Ar/CO2. Temperature was kept above 30oC using a temperature control unit and a hot water coil.

Results and Discussion

Activated sludge with partial nitrification (AS/PN) process was in operation for over 140 days. During this period, different DO monitoring and controlling methods were applied. It was found that the manual control produced best results at DO of around 0.5-0.6 mg/l. During the operation period, steady state was achieved on day 100 150. During this period, COD removal efficiency and concentration in the effluent were averaged at 90% and 63 mg/l respectively (Figure 3). Hence, the effluent has very low organic carbon which is highly suitable for ANAMMOX process. Furthermore, during this period, almost half of ammonium was oxidized and converted to nitrite. Concentrations of both ammonium and nitrite were fluctuated around 100 180 mgN/L (Figure 4). Addition of ammonium chloride was done from day 82 onwards to compensate for the loss in ammonium concentration in the industrial wastewater. A small amount of nitrate was produced; however, the concentration was always less than 30 mg N/L which should not affect ANAMMOX process. In essence, AS/PN process was stable during the experimental period and successfully produced effluent which highly suited for the ANAMMOX process. Key results from AS/PN process is illustrated in Figure 3 and 4, while the key results from the ANAMMOX process is shown in Figure 5. The ANAMMOX pilot scale unit was in operation for 120 days. Initially, enrichment was carried out using synthetic wastewater. On day 89, effluent from AS/PN process was transferred into the reactor. During 30 days period using industrial wastewater, the ANAMMOX process performance was satisfactory. Nitrogen loading and removal rates on day 114 were 0.33 and 0.27 kg N m-3 d-1 respectively. This suggested that AS/PN effluent was suitable for ANAMMOX bacteria and process could be performed successfully. Although, the removal rate was not high as in previous experiments, this could be due to short acclimatization period. Generally, ANAMMOX process requires around 3-9 months to start-up. Another experiment on laboratory scale unit showed that removal rate could be up to 0.6 kg N m-3 d-1 with stability (data not shown).

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COD Inf. 1200 1000

COD_Eff.

% Removal 100 80

COD (mg/L)

800 600 400 200 0 0 20 40 60 80 Time (day) 100 120 140

40 20 0

Figure 3: COD concentrations and removal efficiency from AS/PN process

NH4N Inf. 500 400
Concentration (mg N/ L)

NH4N Eff.

NO2N Eff.

NO3N Eff.

300 200 100 0 1 21 41 61 81 Time (day) 101 121 141

Figure 4: Concentrations for different N species from AS/PN process

NLR 0.35 0.30 0.25 NRR

kg N m -3 d -1

0.20 0.15 0.10 0.05 0.00 0 20 40 60 Time (day) 80 100 120

Figure 5: Nitrogen loading rate (NLR) and removal rate (NRR) for ANAMMOX process
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% Removal

60

Networking

Within ARRPET
The networking group was rationalized and re-arranged to reflect the need of each NRI in this project during the combined regional workshop in Bali (June, 2006). In this respect, AIT has actively carried out collaborative researches including: Microbial population analysis with ARI: AIT has sent Anammox biomass sample to ARI for isolation as well as pure culture development. Furthermore, AIT also sent DNA bands from partial nitrification and Anammox reactors to ARI for sequencing. The results from this research will be published under a joint publication. Biofloc/Bilfilm development and characterization with IIT-B: AIT and IIT-B has set up a collaborative research on granule characterization of the Anammox inhibited biomass. Preliminary experiments were carried out at AIT, and samples were sent to IIT-B for image analysis to determine floc size distribution. The result has shown great variation between normal and inhibited aggregate size. Thus, further investigation is necessary and will be conducted within the next 6 months. Microbial analysis and pilot plant investigation with KMUTT: KMUTT has assisted AIT in setting up of Fluorescence In Situ Hybridization (FISH) laboratory procedure as well as providing connection to the suitable seafood industry.

Outside ARRPET
AIT has undertaken a pilot scale investigation on membrane bioreactor (MBR) with an acrylic fiber industry in Thailand. The collaborative researches include from cleaner production with a focus on reuse and recycle of treated effluent. Furthermore, investigation of pilot scale unit membrane filtration and MBR was also conducted at the industry. Results from the studies have identified various options on reusing of effluent and ways to enhance COD removal efficiency of the existing treatment plant. As a result, the industry decided to install a membrane filtration unit to reuse treated effluent from the wastewater treatment plant as a make up water in the cooling towers. DISSEMINATION

Workshops Organized:
Table 1: List of workshop organized by AIT-WWTM Networking activity Training on UASB and ANAMMOX Process Training Workshop Techniques
WWTM Project

Remarks AIT, Thailand; 21-25 March 2005 AIT, Thailand; 21-25 March 2005 ARRPET Phase II

on

Molecular

12

Publications (other than research publication):

Annachhatre, A.P. (2005). Advanced, Appropriate and Affordable Technologies for Domestic Wastewater Treatment and Reuse in Tropical Region, Proceedings of UNESCO Workshop on Integrated Urban Water Management on Humid Tropics, Iguassu Falls, Brazil, 2-3 April 2005. Khin, T. and Annachhatre, A.P. (2004) Novel Nitrogen Removal Processes presented at the First Thailand-Korea International Symposium on Environmental Remediation held at AIT on 20 August 2004. San Francisco, L.S. and Annachhatre, A.P. (2004). Sustainability of water resources. IWA International Conference, Western Australia, November 2002., 167-174. Sinha, B. and Annachhatre, A. P. (2005). Inhibition of Nitrite Oxidation during Nitrification. ARRPET Newsletter August 2005. Lamsam, A. and Annachhatre, A. P. (2005). Cultivation of Anammox Bacteria: Tricks or Treats. ARRPET Newsletter December 2005. CAPACITY BUILDING

Man- Power:
Miss Le Tuyet Minh - Master student (completed) Miss Mayuree Tonkham Master Student (completed) Mr. Bhaskay Ray Master Student (completed) Mr. Kawin Ruamsuke Master student (completed) Mr. Tharmalingam Suthakar Master Student (completed) Mr. Apipong Lamsam PhD Student (on-going) Mrs. Banashri Sinha - PhD Student (on-going) Miss Sawanya Laohaoprapanon Master Student (on-going) Mr. Nguyen Phuc Thanh Master Student (on-going) Mr. Nuttapol Tanadehangsaeng Master Student (on-going) Mr. Phrompol Chantrasakdakul Master Student (on-going) Ms. Raquel P. Pedrajas Senior Research Associate

Laboratory Infrastructure:
Major equipment (>50,000 SEK) Microscope with fluorescence, Program Meta System (Isis FISH) Minor equipment (< 50,000 SEK) Datalogger ML22, Anammox Gas-lift Reactor (consisting of reactor height 2175 mm, riser height 1700 mm, body made of stainless steel 304 thickness 1.2 mm, cover made of stainless steel 304 thickness 2 mm, steel plate thickness 6 mm, stainless valve 1/4", steel frame, height 200 mm width 80 mm). Pilot scale experimental setup consisting of Annamox Gas-lift Reactor, Heater (HCL19X300-220V-1000W), Temperature sensor (FWP-19A-6.35X100-3M (RTD Pt 100 DN), Drive MFLEX, L/S 600 RPM, 115/230V, Pumphead, L/S, Easy-Load 3, SS Maintenance Kit for Do Sensor (Model: ZBK 600), WTW pH Sensor (Sensolyt

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ECA), O2 4050e wall O2 electrode model, InPro 6050/120, Inlab 413-SG pH probe for Seven-Go pH meter, Computer. Wall Cabinet, Desk, pH-DO Portable sensor, Micropipette, Incubation oven, Hot Plate, Dispensers (2.0-50.0 mL), Partition dark room for FISH analysis, pH meter, Computers, Printer, UPS, Masterflex pump and accessories, pH Probe, Automatic Burettes. Open Issues Unresolved issues from ARRPET II experiments are as follows: Dissolved oxygen (DO) control in AS/PN process may require optimization using automatic control to enable full scale application. Combined AS/PN-ANAMMOX should be tested with an industrial wastewater following a UASB reactor. Furthermore, experiments should be conducted to optimize both processes under real situation of influent flocculation and varying temperature in a tropical climate. Maximum capacity of ANAMMOX process in the gas-lift reactor should also be verified. Detailed cost-benefit analysis and EIA studies. Characterization of microbial community from efficient AS/PN and ANAMMOX processes should be conducted.

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CENTEMA Report: Sustainable Development of Tapioca Processing Industry in Vietnam

Contact Person(s): NGUYEN TRUNG VIET, TRAN THI MY DIEU, HUYNH NGOC PHUONG MAI
CENTER FOR ENVIRONMENTAL TECHNOLOGY AND MANAGEMENT VAN LANG UNIVERSITY C4/5-6 DINH BO LINH, BINH THANH DIST., HCMC VIETNAM e-mail: centema1@hcn.vnn.vn 1. TECHNOLOGY DEVELOPED THROUGH RESEARCH

The rapid growth of industrialization and urbanization in Vietnam is putting severe stress on natural resources and on the environment, forcing the country to face a number of serious environmental problems, such as water and air pollution, degradation of land resources, soil erosion, over-exploitation of natural resources, and threats to the ecosystem. Important reasons for the rapid increase in environmental pollution are mismanagement and limitations in the prevailing level of technology applied in industrial production processes and in waste treatment. Tapioca processing industry is chosen as a case study for our research. Environmental problems from the tapioca processing industry can be divided into specific categories as will be presented below. Resource consumption: water consumption for starch extraction process and energy consumption (coal, petroleum, gas or electricity) for starch drying. Wastewater: in order to produce 1 ton of starch, a tapioca processing factory discharges a huge amount (i.e. 15-20 m3) of wastewater, which contents high organic matters, nutrients, cyanide concentration and very low pH. This is cause serious problem for the surface and underground water quality in local area. Solid waste: including root skins and fibrous residues, these may become local source of malignant and obnoxious odors during the drying and storing. A portion of the solid waste is dumped may become environmental pollution. Air pollution: mainly originates from the combustion of fuel (dust, CO, CO2, SO2, NO2) and comes from the greenhouse gases released from the degradation of wastewater in anaerobic ponds and from solid waste. Another kind of air pollution is obnoxious odors from degradation of organic matter. The surveys on tapioca production processes showed that there are several possibilities to reduce and reuse non-products (including byproducts and wastes) from tapioca processing units. Based on that, it is possible to reduce and eliminate environmental impacts as well as to improve production efficiencies of tapioca processing units. Several international literatures show that there are many different ways to get rid of wastes and achieve zero (or close to zero) waste industrial systems. Certainly, each approach has its strengths and weaknesses and its application often works under specific conditions. Integration of different approaches can overcome the 15 ARRPET Phase II

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weaknesses and shortcomings of individual approaches. The starting point to develop such a methodology is formed by the material and energy flows in industrial systems. This aims at developing a systematic methodology on how to analyze various options to approach a zero waste industrial ecosystem by prevention of waste, minimization of waste, reuse and recycling within one company and within a wider network of companies and waste treatment option. Then, the objective of ARRPET phase 2research is to develop a general model of zero waste industrial ecosystem for tapioca processing industry in Vietnam. To achieve the objective, CENTEMA carried-out the research step by step: Primary surveys and data collection is carried out at 6 tapioca processing villages and 3 large-scale tapioca processing companies. Detailed study is carried out for 2 selected companies. Data from detailed studies to develop the physical-technological model relate mainly to material and energy flows within research units and related components of the model. Within research units the date of input (raw materials, chemicals, energy, water supply, labor) and output (main products, solid waste, byproduct, wastewater, air pollutants) are collected and studied based on material balances. Besides, the data of related components (wastewater treatment plant, fish culture, cassava culture, livestock feed production, composting plant, biogas plant) also are collected. Based on the data, the research is carried out and conducted: Development a methodology Assess strengths and weakness of pollution prevention approaches including endof-pipe treatment, waste reuse and recycling/waste minimization, cleaner production and industrial ecology; Utilize strengths of these pollution prevention approaches to develop a method to move an existing industrial system toward a zero waste industrial ecosystem (physical-technological model development); Apply a triad-network of Ecological Modernization Theory to push a physicaltechnological model of developed zero-waste industrial ecosystem from table to work.

Development an eco-industrial system Calculate material balances within tapioca processing factories using the results of detailed studies; Ascertain waste exchange ability oriented toward eco-industrial system (closed and/or open eco-industrial system); Material balances among tapioca processing factories and other components of industrial ecosystem; Designing physical-technological industrial ecosystems (closed and/or open industrial ecosystems) including: tapioca processing factory, composting, fish culture, livestock feed production, cassava cultivation, wastewater treatment system. Operation (pilot-scale model) including: wastewater treatment system, fish culture, composting plant, plan cultivation.

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Policy and regulation including: Collection and evaluation of existing environmental policy and regulation Preparation of new policy to support the eco-industrial system.
Tapiocaproduction process
Watersupply

Cassava harvesting

Solid wastes Solid wastes

Gas recovery Gas recovery

Wastewater Wastewater

Market

Cattle -food processing

Compos ting

Waste sludge

WWTS

Cattle Breeding

Fish culturing

Receiving source

Cassava culturing

Fig. 1 Recommended sustainable development for tapioca processing industry.

Based on the results of our research, we can recommend an almost perfect system for the tapioca industry for Vietnam. It combines pollution prevention with energy recovery and resource protection. With the proposed integrated treatment system, which incorporates waste minimization and reuse of valuable compounds from the waste, a sustainable development of the tapioca industry can be achieved. Waste and wastewater can therefore really comprise a resource instead of a headache. The treated wastewater can be reused in aquaculture or for irrigation in the area. Peel waste can be used for composting, and the compost produced can be applied for the cassava cultivation or other industrial crops. Fibrous residues can be used for animal feed production. In this way, a zero waste industrial ecosystem can be created (see Fig. 1). This industrial ecosystem can be applied in case of tapioca processing villages, which solid waste and wastewater can be gathered and treated at the central treatment units in the area.

List of research publications during ARRPET phase 2 (2004 2007)

a. International conference: Mai, H.N.P., Duong, H.T., Trang T.T.T. and Viet, N.T. (2004), Sustainable Treatment of Tapioca Processing Wastewater in South Viet Nam. In proceedings: International Conference on Wastewater Treatment for Nutrient Removal and Reuse 2004, Asian Institute of Technology Thailand, 26-29 January 2004, Vol 2, pp. 373-384. Mai, H.N.P., Duong, H.T., Trang, T.T.T. and Viet, N.T. (2004), UASB Treatment of Tapioca Processing Wastewater In South Vietnam. In proceedings: International Conference on 10th

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World Congress Anaerobic Digestion 2004 29th Aug. to 2nd Sep, 2004 Montreal, Canada, Vol 4, pp. 2420-2424. Kragic, D., van Buuren, J.C.L. and Trang, T.T.T (2005). Evaluation of Industrial Wastewater Treatment in Ho Chi Minh City and the Southern Provinces of Vietnam. In proceedings: International Conference on Environmental Management of Urban and Industrial Infrastructures in Asia. HCMC, Vietnam, 11-12 November, 2005. b. International papers: A.P.J. Mol and Tran Thi My Dieu (2006), Analysing and Governing Environmental Flows: the Case of Tra Co Village, Vietnam, NJAS The Wageningen Experience with Research and Education for Development- Wageningen Journal of Life Sciences, Volume 53, Number 3-4, pp. 301-317. Dieu, T.T.M. (2006), Greening Food Processing Industries in Vietnam: Opportunities and Constraints, Environmental, Development and Sustainability (2006): 8: 229-249, Springer 2006. c. Book chapters: Mai, H.N.P. (2006), Integrated treatment of tapioca processing industrial wastewater Based on environmental bio-technology. PhD dissertation, Wageningen University. The Netherlands. 2. PILOT SCALE APPLICATION Wastewater treatment plant

OPTION

Biogas

Bar Screen
WASTEWATER

Grit Chamber

Skimming & Settling

Neutralization Flow Neutralization Equalization

Animal feed

Sludge

Final settling Stabilization pond

EFFLUENT

Activated sludge

Stabilization pond

Air blower Waste sludge Return sludge

Fig. 2 Technology for treatment of tapioca processing wastewater, capacity 10m3/d.

A pilot-plant scale version for wastewater treatment with a capacity of 10 m3/d has already been designed and installed (see Fig. 2). The system start-up was commenced in the middle of December 2005 at KMC Tapioca Starch Factory, Binh Phuoc 18 ARRPET Phase II

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Province. The performance data collection show that it is possible to apply the technology which aerobic combined anaerobic process for tapioca wastewater with very high treatment efficiency and the biogas production fluctuates from 320-350 liter per kilogram COD removal. A successful application of this treatment system can contribute significantly to a further sustainable development of the tapioca processing industry in Vietnam by minimizing the environmental pollution. Based on the results of pilot-scale operation, the estimate shows that the required treatment system with a capacity of 2,000 m3/d will produce 4,200 6,200 m3 biogas per day (>60 % methane gas), which is equivalent to 2,600 3,800 liters of FO per day. And energy cost savings to the plant are estimated at 850 1,260 US$ per day. If 1 m3 biogas (with 60% methane) is equal to 1.7 kWh of electricity (data from the power plant using biogas of Go Cat landfill, HCMC, Vietnam), then the power plant can generate 7,100 10,500 kWh per day. The proposed wastewater treatment system combined with energy recovery can also benefit from international carbon trading and help support financing projects through the Clean Development Mechanism (CDM). In this project based on converting wastewater to energy and profit - both the technology and the business model are highly possibility. It also shows how strong returns can be generated for investors using CDM. Besides, for the solid waste treatment, a composting plant and cassava cultivation are carried out also at pilot-plant scale. Composting plant The materials consisting of cassava peel root and activated sludge from the tapioca wastewater treatment system used for the input of composting experiments. The composting method and technical process of composting are presented as Table 1 and Fig.3 below.
Cassava peel

Cutting

Mixing with Sludge EM - Mixing - Water - EM i

Screening

Aerobic composting

Curing

Screening
Mixing with micro nutrients etc. (if applicable)

Residue

COMPOST

Fig. 3 Technical process of composting

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Table 1 Introduction of pilot scale of composting experiments

Stage 1 Experiment Composting method Material C/N Blending ratio (peel root : sludge) Initial weight of the pile (kg) Experiment Composting method Material C/N Blending ratio (peel root : sludge) Initial weight of the pile (kg) 1 Aerated static pile 2 Aerated static pile 3 Periodic turning pile Peel root + sludge 25 3:1 139 3 Periodic turning pile Peel root + sludge 25 1,6:1 200 4 Periodic turning pile Peel root 64 144 4 Periodic turning pile Peel root + sludge 25 1,6:1 200

Peel root + sludge Peel root 25 64 3:1 139 144 Stage 2 1 2 Periodic turning Periodic turning pile pile Peel root + sludge Peel root + sludge 30 30 2,25:1 2,25:1 200 200

After about 35 days of composting and 10 days for curing, the final composting products were applied for cultivation. The chosen plants for cultivation are corn and cassava presented in following part. Cultivation of plant The corn and cassava were chosen as the objects of study on compost application in cultivation. Based on the process showing in Fig.4, the pilot scale of corn cultivation experiments and cassava cultivation experiments were operated and introduced.
Land/young tree preparation Fertilizing by compost Planting/ observation Harvesting

Fig. 4 Compost application process for cultivation. Table 2 Introduction of pilot-scale corn cultivation experiments Experiment Type of plant Area (m2) Density of plant (trees/m2) Number of plant 1 Fertilizing by compost from cassava hard roots & wood shells Corn 8 m x 9 m = 72 m2 4 288 2 Fertilizing by sludge from stabilization pond 1 Corn 8 m x 12 m = 96 m2 4 384

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Table 3 Introduction of pilot-scale cassava cultivation experiments Experiment 1 2 Fertilizing by compost Fertilizing by sludge from from cassava peel & stabilization pond sludge Cassava 8 m x10 m = 80 m2 2 160 Cassava 8 m x 10 m = 80 m2 2 160 3 No fertilizing

Type of plant Area (m2) Density of plant (trees/m2) Number of plant (trees)

Cassava 8 m x 10 m = 80 m2 2 160

Key results of pilot-scale application The UASB reactor system combined with an aerobic activated sludge process and stabilization ponds, represents a superior solution for the treatment of highstrength wastewater like tapioca-processing wastewater under Vietnamese conditions, because it enables meeting the severe Vietnamese effluent discharge standards; The treated wastewater can be reused for irrigation in the area of cassava cultivation or for agriculture purpose; The proposed wastewater treatment system combined with energy recovery can benefit from energy cost saving and CDM program. This is the persuasive reasons for industries and financial investors to implement this treatment technology in Vietnam. Sustainable development can be achieved with waste minimization and reuse valuable compounds from waste; To apply the sustainable development for tapioca industry, the study on the policy/regulation for technical assistance and financial assistance for the factories need to be carried-out; The wasted sludge from wastewater treatment plant and wasted root peel can be reused as the material for composting; The compost produced is satisfied Vietnamese standard for compost product and it can be applied for the cassava cultivation or other industrial crops.

3. NETWORKING Within ARRPET Networking activity AIT Description CENTEMA attended the training workshop in AIT on: UASB and ANAMMOX Processes during 21-25 March, 2005. Molecular techniques on ANAMMOX bacteria during 21-25 March, 2005. KMUTT Technology exchange on from 27-28 June, 2005; The Sustainable Shrimp Farming Workshop

Collaboration in holding the Sustainable Aquaculture Engineering Approach Workshop from 8 9 February, 2007 in Hua Hin, Thailand.

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IITB

Research on granulation of sludge from UASB in pilot scale; Information exchange about agro-industry; Technology exchange about Advanced Analytical Techniques (attending the Advanced Analytical Techniques workshop during 2-4 April, 2007).

News letter Outside ARRPET Networking activity

Responsible for ISSUE 3

Description Collaboration in carrying-out the research. Collaboration in carrying-out the research. Supporting the investigation of tapioca processing industry in village and company. Co-hosting the workshop on Activated Sludge Process operation and maintenance during 5 -17 July 2004. Creating the good condition and extremely support for operating the wastewater treatment system, pilot of composting as well as the cultivation. Cooperation with CENTEMA in holding the workshop on Appropriate Criteria for Assessing the Performance of Waste Treatment Systems in September 30, 2005 in HCMC, Vietnam.

Dept. of Environmental Technology and Management Van Lang University Van Lang Company Department of Natural Resource and Environment of Tay Ninh province and Binh Phuoc Province Division of Solid Waste Management DONRE - HCMC KMC tapioca starch Co., Ltd. Vietnam

Vietnam Environment Protection Agency

4. DISSEMINATION Workshop No. Workshop Within ARRPET 1 Training course: UASB operation and maintenance 2 Training course: Activated Sludge Process operation and maintenance 3 Training course: The technical skill for laboratory staffs 4 National workshop: Appropriate Criteria for Assessing the Performance of Waste Treatment Systems 5 National workshop: Development of Eco-industrial Park: A Case study of Linh Trung Export Processing Zone (supported by Department of Science and Technology, HCMC) 6 Sustainable Aquaculture Engineering Approach (cooperation with KMUTT) Venue CENTEMA - Vietnam CENTEMA - Vietnam CENTEMA - Vietnam HCMC, Vietnam Department of Science and Technology, HCMC, Vietnam Hua Hin, Thailand 8 9 February, 2007 Duration 7-19 June 2004 5-17 July 2004 13-24 December 2004 September 30th, 2005 July 26th, 2006

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Outside ARRPET 7 Waste Management (supported by DAADGermany) 8 Sludge Management in HCMC (supported by Vietnam Australia Environment Joint Stock Co.) Publication No. 1.

Van Lang University, HCMC, Vietnam HCMC, Vietnam

14-16 December, 2006 23 24 April, 2007

Articles Ecological Transformation Of The Tapioca Processing Industry In Vietnam

Le Van Khoa* and Tran Thi My Dieu** * DONRE ** CENTEMA, Van Lang University and Environmental Policy Group, Wageningen University
Greening Industrialization in Asian Transitional Economies, Edited by Arthur P. J. Mol and Joost C. L. van Buuren, Lexington Books Publisher, pp. 199-224.

Newsletter No.3

2.

Environmental Reform Of Food-Processing Industry: An Industrial Ecology Approach on Bien Hoa 1 Industrial Zone, Vietnam
Tran Thi My Dieu*, Arthur P. J. Mol** and Wim H. Rulkens***
*CENTEMA, Van Lang University and Environmental Policy Group, Wageningen University ** Environmental Policy Group, Wageningen University ***Environmental Technology Group, Wageningen University International Journal of Business and Society, Volume 5, Number 1, January 2004, Sarawak, Malaysia, pp. 29-57.

Newsletter No.3

3.

Integrated Treatment Of Tapioca Processing Industrial Wastewater - Based On Environmental Bio-Technology

Huynh Ngoc Phuong Mai
Center for Environmental Technology and Management, C4/5-6 Dinh Bo Linh, Binh Thanh Dist., HCMC, Vietnam. Extracted from PhD thesis Integrated treatment of tapioca processing industrial wastewater - based on environmental bio-technology.

Newsletter No.4

Technical report and book chapter No. 1. 2. 3. 4. Technical report within ARRPET phase 2 Model of tapioca processing industrial ecosystem at village level: A Case of Tra Co Tapioca Producing Village (extracted from the appendix of the project progress report in August 2005). Sustainable development of tapioca processing industry: Methodology and Case Studies (extracted from the appendix of the project annual report in February 2006). EIA report for the project of sustainable development of tapioca processing industry in Vietnam (extracted from the appendix of the project annual report in February 2006). Cost benefits analysis report for sustainable development of tapioca processing industry in Vietnam

5. CAPACITY BUIDING Within the research, the manpower is reinforced by training and improving for the field of environmental protection and sustainable development, including: 01 PhD student, 05 students and 08 research staffs. Minor equipments have been purchased during period:

+ For field survey: pH meter, DO meter, gas meter. 23 ARRPET Phase II

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+ For the laboratory: oven, technical balance, distillation apparatus for cyanide, composting reactors, cutting machine for composting.

6. OPEN ISSUES

Results from the research shows that it is possible and good idea to develop a zero waste emission eco-industrial for tapioca processing industries. This is an innovative strategy for sustainable development of tapioca processing industry by reducing waste, reducing the consumption of resources and increasing the recycling of materials and energy. However how to put the theoretical model develops from this research to work is still a question. How to advise the tapioca factories put into their plant and apply solution from this research to their own factories. Finally, how to deploy this model for the whole country? This oriented questions show the needs of further research to establish so-called guideline to develop a tapioca eco-industrial system in general full scale apply.

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IITK Report: Domestic Wastewater Treatment in India: Optimization of UASB Reactor

Contact Person: Saumyen Guha Department of Civil Engineering Indian Institute of Technology Kanpur, UP 208016, INDIA e-mail: sguha@iitk.ac.in 1. Technology Developed through Laboratory Research

The advent of high rate anaerobic technology and its proven applicability for the treatment of high strength wastewater has lead to attempts towards its implementation for domestic sewage and low-strength wastewater in the countries with warmer climates. The UASB process, on account of their low installation cost and low operation and maintenance overheads, has been installed in some locations in the developing countries, such as, India, Colombia and Brazil etc. The UASB process relies on anaerobic degradation of organic wastes by combining the advantage of high active biomass retention with high void volume. Anaerobic microbial granulation, considered as a key parameter for successful operation and stable performance of an UASB reactor, is a natural process under favorable conditions as a result of tendency of bacteria to self immobilize. The UASB process for the treatment of low strength wastewater, however, suffers from the following shortcomings: longer start-up times, inability to form self-immobilized bacterial granules and very low bio-gas recovery. Primary objective of the project at IITK was to achieve better granulation in UASB reactors treating low-strength wastewater, by maintaining appropriate microbial ecology as well as physico-chemical environment inside the reactors. In the literature, many natural and synthetic polymers were reported to enhance granulation but the reports once again, were mostly limited to high strength wastewater. The choice of an additive in the ARRPET project was governed by scientific understanding of the process of granulation, low cost and local availability. The Reetha seed (Sapindus trifoliata) extract was selected as a plant polymer additive. Reetha is cheap and easily available in the region and contains high amounts of anionic, cationic and non-ionic polymers. For comparison, we chose Chitosan a cheap water-absorbing polymer derived from the shells of the crustaceans. Five laboratory scale UASB reactors were run in parallel with same flow rate, influent and environmental conditions. Four of these reactors were augmented with the following additives: (i) Bulk Reetha seed extract, (ii) Cationic fraction of Reetha seed extract, (iii) Anionic fraction of Reetha seed extract and (iv) Chitosan. No additive was added in the fifth reactor and it served as the control. Influent and effluent samples from each reactor were analyzed as follows: for total COD and filtered COD every alternate days; for pH and alkalinity twice a week; TSS, VSS and VFA twice a week. Production of methane and total gas were monitored twice a week. Sludge 25 ARRPET Phase II

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samples form the reactors were analyzed for TSS, VSS once a week. Extra-cellular polymers (ECP) and its component sugar, lipids, and protein were analyzed at approximately 20 days interval. Total and Acetoclastic methanogenic activity tests were conducted once every month. While reactor operation and analyses listed so far were conducted at IIT Kanpur, a close collaboration was maintained with the NRI of IIT Bombay throughout the study period for granule examination. Sludge samples were fixed and sent to IIT Bombay at a frequency of 15-20 days. They provided us with the image analysis data showing size distribution. For some select samples, granules were also examined under Environmental Scanning Electron Microscope (ESEM) and Transmission Electron Microscope (TEM). Initial attempt during the Phase I of ARRPET to enhance granulation using alum as coagulant failed and the aluminum was found toxic to the microbial population. Thereafter, enhancement in granulation using the natural polymers was achieved in Phase I of ARRPET using synthetic wastewater with an influent COD of 800 mg/L. These two experiences helped us in understanding the mechanism of granulation in the treatment of low strength wastewater. In Phase II of the ARRPET, the larger granules generated with natural polymer additives were first tested for the stability by subjecting them to wide variations in flowrate and COD concentration. The large granules were stable up to influent COD concentration of 250 mg/L and flow rate variation of 0.5-2 times the design flow. The long term reactor performance and stability of the granules were assessed by running these reactors for 900 days. Based on the understanding of the process of granulation using synthetic wastewater, the enhanced granulation experiments were designed with real domestic wastewater with influent COD concentration varying in the range of 90-200 mg/L during a 24 hr period and COD/SO42- ratio of < 2. Enhancement of granulation was once again observed. One key factor in all these studies has been that the seed sludge was dispersed and did not contain any granules. However, all the reactors reached steady state in ~60 days. With real domestic wastewater, up to 80% COD removal was obtained with only 2.6 hr HRT in the sludge zone and an overall HRT of 4.5 hr. Although, the influent COD and the resulting OLR shows large (up to 150%) diurnal variation, the effluent COD has a variation of only 10%. At present, the reactors with real domestic wastewater have completed operation for 700 days and the granules continue to grow. The granules of size up to 3-4 mm have been observed and nearly 60-80% of the sludge bed has undergone granulation leaving very little dispersed sludge in the bed. Such large sizes of granules with low influent COD concentration in the range of 90-200 mg/L have not been reported. Thus, the additives helped to maintain larger granules in the sludge bed. This allowed higher flow rate and increased resilience to shock loading. We have been able to demonstrate that the protein fraction of the exo-cellular polymeric substances contributes to the structural integrity of the granules and that the level of total exo-cellular polymeric substances does not reflect adequately the state of granules. Domestic wastewater often contain COD/SO42- ratio of < 2. According to literature, this leads to interference by sulfate reducer and cause reactor failure. We have not observed any such interference so far although the condition is met. Initial observations indicate that this may have been averted in our reactor by putting a selection pressure on the sulfate reducers at the initial stages. At present, efforts are on to confirm and standardize this condition such that UASB reactor can be started with unfavourable COD/SO42- ratio.

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During the third year of Phase II, a cost benefit analysis was conducted for the technology developed using the activated sludge process as the benchmark. The technology developed in ARRPET was found to be economically beneficial and environment friendly. The experience of operating UASB reactors and many other biological processes in the ARRPET project showed the importance of monitoring the microbial ecology. This may indicate crucial stages of the reactor performance as well as impending reactor failure. We made an effort to devise a simple process based on analysis of phospholipid fatty acids. The method is appealing because the analysis can be completed in a few hours and reliable estimates are possible. This will enable quick intervention to reactor operation, if required. Although the PLFA analysis are well established in the literature, lack of standard deconvolution strategies make them unusable for routine application to reactors. We have devised and tested a novel apriori approach to deconvolution of the PLFA signature of mixed culture. The efforts are on to apply it for the routine monitoring of UASB reactor. However, some more problems remain such as, modification of PLFA-FAME analysis for isoprenoids present in archea, standardized posteriori deconvolution approach, etc. For the minor issue project, we had demonstrated that it is possible to degrade the azo dyes using fixed bed fungal column containing Phenerocheate Chrysporium. Publications from the project so far are as follows: Journals (peer reviewed) Dey, D. K.; Guha, S. (2007) Determination of Community Structure through Deconvolution of PLFA-FAME Signature of Mixed Population, Biotecnology Bioengineering, V. 96, No. 3, pp409-420. Tiwari, M. K.; Guha, S.; Harendranath, C. S.; Tripathi, S. (2006) Influence of Extrinsic Factors on Granulation in UASB Reactor, Applied Microbiology and Biotecnology, V. 71, No. 2, pp 145- 154. Tiwari, M. K.; Guha, S.; Harendranath, C. S.; Tripathi, S. (2005) Enhanced granulation by Natural Ionic Polymer Additives in UASB reactor treating lowstrength wastewater, Water Research, V. 39, pp 3801-3810. Tiwari, M. K.; Guha, S.; Harendranath, C. S. (2004) Enhanced granulation in UASB reactor treating low-strength wastewater by natural polymers, Water Science and Technology, V. 50, pp 235-240. Sondhi, A.; Guha, S.; Harendranath, C. S. and Singh, A. Effect of Aluminum on Granulation in UASB Reactor, in preparation. Singh, R.; Quaff, A. R.; Guha, S. and Harendranath, C. S. Stability of Granules in UASB Reactor Treating Low Strength Wastewater: Flow Variability and Low COD, in review. Quaff, A. R.; Arockia, V.; Guha, S. and Harendranath, C. S. Population Shift During Start-up of UASB Reactor Treating Domestic Wastewater, in preparation.

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Conferences Singh, R., Quaff, A. R., Guha, S., Harendranath, C. S. Stability of Granules in UASB Reactors Treating Low Strength Wastewater: Flow Variability and Low COD, International Conference on Civil Engineering in New Millennium, CENEM 2007, Jan 11-14, 2007, Bengal Engineering and Science University, Shibpur, Howrah, West Bengal, India. Quaff, A. R., Arokia, V. J., Guha, S., Harendranath, C. S. Population Shift During Startup of UASB Reactor treating Domestic Wastewater, International Conference on Civil Engineering in New Millennium, CENEM 2007, Jan 11-14, 2007, Bengal Engineering and Science University, Shibpur, Howrah, West Bengal, India. Quaff, A. R., Arokia, V. J., Singh, R., Guha, S., Harendranath, C. S. Startup of UASB reactor using low strength domestic wastewater, International Workshop on Biotechnology of Anaerobic Bacteria & Archaea, March 3 & 4, 2006, Agharkar Research Institute, Pune, India. Arokia, V. J., Quaff, A. R., Singh, R., Harendranath, C. S., Guha, S. Application of Image Analysis and Electron Optical Techniques in the Evaluation of Anaerobic Process, International Workshop on Biotechnology of Anaerobic Bacteria & Archaea, March 3 & 4, 2006, Agharkar Research Institute, Pune, India. Singh, A, Shetye, B, Guha, S., Harendranath, C. S. Application of ESEM to study granulation in an UASB reactor treating wastewater, XXVI Annual Conference on Electron Microscopy and Allied Fields, organized by Electron Microscope Society of India at CPRI, Shimla during April 16-18th, 2003 (Best poster Award). 2. Pilot Scale Application

In India, the domestic wastewater treatment is public sector unit and there is no funding for testing pilot plants. However, the private initiatives have many units which treat composite wastewater from domestic sources and small scale industries. Vapi Waste and Effluent Management Co. Ltd. (VWEMC Ltd.) at Vapi, Gujarat is one such organization. They agreed to built, operate, maintain and monitor a pilot scale reactor based on our design and operation instruction. A 100 cu. m./d UASB pilot plant was constructed with the joint collaboration of IIT Kanpur, IIT Bombay and VWEMC Ltd. The plant was commissioned in July 2006 to treat composite wastewater from various industries such as Chemicals, Dyes, Intermediates, Pharmaceuticals, Paper & Paper Board, Cosmetics, Pigments, Paints, Textile, Glass, Engineering etc. mixed with domestic sewage from the twon of Vapi. This mixed wastewater has COD/SO42- ~ 1 with COD concentration varying between 800-2200 mg/L and 5-day BOD varying between 100-200 mg/L. Wastewater from the equalization tank directly fed to the UASB reactor without any pretreatment. The reactor was seeded with a mixture of sludge from the dyring bed and fresh activated sludge. The mixing ratio was selected based on VSS/TSS ratio and methanogenic activity of the sludge. The seed sludge was flocculant in nature and did not contain any granules. Chitosan was used as additive for enhancement of granulation. First dose was added at the startup and second dose was added on 170th day of operation.

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The plant at present is removing 20 % COD and 40-50 % BOD with a total HRT of 10 hr. It may be noted that the influent contains difficult to degrade substrates from the industrial effluent. A standard UASB reactor operating for more than a year with 19 hr HRT and effluent recycle gives inferior performance compared to the ARRPET reactor. The pH and VFA level have stabilized within 90 days. The VSS/TSS of sludge has also stabilized at 0.5. The color removal is 30% and TSS removal is 40%. Continuously increasing Methanogenic activity shows that the Methanogens are establishing in the reactor and the gas production has started at 94 L/day. A TEM result shows that Methanosarcina has established in the reactor. The exocellular polymeric substances are increasing with its protein fraction indicating healthy granules. Granules size is increasing continuously. Particles greater than 0.1 mm stands at 70% and are growing larger. The particles greater than 0.3 mm are about 21 % and increasing. The VFA level and conversion of the harder to degrade substrates to easily degradable substrates shows significant scope of improvement and need for research in that direction. Considering that 87 such combined effluent treatment plants (CETPs) are already operational in the country and the government subsidy towards setting up more such plants, there is a need to evolve an appropriate technology in the country for treating composite wastewater. At this point, plan is to demonstrate the technology in the pilot plant for pure domestic wastewater and consider the problem of the composite wastewater in Phase III of ARRPET. 3. Networking

One of the key goal of the ARRPET Phase II was to form close network amongst the NRIs and solve the mutual problems through networked research. We maintained a close collaboration with the NRI at IIT Bombay for the granule characterization of the UASB reactors. The effort yielded many a fruitful insight into the processes. With the help of transmission electron micrographs from IIT Bombay and the biochemical parameters measured at IIT Kanpur, we were able to notice a change in microbial ecology during the first 100 days of the start of the UASB reactors with real domestic wastewater. The microbial ecological shift was crucial for start of the formation of the granules. A networking with ARI Pune and IIT Bombay are on to establish detailed microbial ecology of the UASB reactor using genetic methods such as DGGE, PCR, FISH. This will then be compared with the results obtained using PLFA based method designed at IIT Kanpur. We also had a networking with the NRI from UoM, Sri Lanka. They were studying metal uptake in the water hyacynth. We at IIT Kanpur were able to isolate and characterize the phytochelatins responsible for metal binding in water hyacynth plant. With the help of IITB, we were able to locate the metal chelated phytochelatins in various parts of the plant. We demonstrated that the metal is initially bound by phytochelatins of smaller molecular weight and later transferred to larger molecules. Pilot plant is being operated in collaboration between IITK, IITB and VWEMCL.

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4.

Dissemination

In addition to the technical publications in peer-reviewed journals, international conferences and national conferences listed in item 1, we also organized workshops, delivered invited lectures and contributed to the newsletter. Some are listed below: Combined Regional Workshop of WWTM and HWTM was held on 11-12th March, 2005. A Report is available at, http://www.arrpet.ait.ac.th/wwtm/newsletter1/index.html. Delivered invited lecture at the training workshops at ARI, Pune (IHWTM NRI) and IIT Bombay. A special training workshop on Statistical Data Analysis for the NRIs of IHWTM and WWTM to be held in 2007. Close contact is being maintained with industries in Vapi, state pollution control board and Central Pollution Control Board so that the developed technology in ARRPET remains available to the stake holders. 5. Capacity Building

Present Project Staff:

Abdur Rahman Quaff (Ph.D. Student) Richa Parasher (Project Associate) Anoop Agnihotri (Project Assistant)
Completed M.Tech. Theses (Available online through IITK Library):

Patel, N. K. (2006). Characterization of Phytochelatins Produced by Water Hyacinth, M.Tech. Dissertation, IIT Kanpur. Dey, D. K. (2005). Determination of Community Structure through Deconvolution of PLFA-FAME Signature of Mixed Population, M.Tech. Dissertation, IIT Kanpur. Singh, R. (2004). Effect of Variable Flow Rate and Low COD on the Stability of Granules in UASB Reactor, M.Tech. Dissertation, IIT, Kanpur. Tiwari, M. K. (2003). Enhancement of Granulation in UASB Reactor by Natural Polymer Additives, M.Tech. Dissertation, IIT, Kanpur. Sondhi, A. (2002). Effect of Aluminum Chloride on the Performance of UASB Reactors Treating Low Strength Wastewater, M.Tech. Dissertation, IIT, Kanpur.
Equipments Procured from ARRPET II fund:

Hereus Biofuge Stratos R, Coy Laboratories Anaerobic Glove Box, Laminar Flow Hood, PC. 6. Open Issues

The following are the open issues from ARRPET Phase II:

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Deveopment of appropriate technology for the treatment of composite wastewater: The CETP concept is widely given preference by the governmental agencies. There are 96 CETPs already established and the number is increasing every year. Majority of the plants are based on power consuming activated sludge process that generates a large amount of toxic sludge. Development of anaerobic technology in this direction will not only cut down on power consumption but also recover valuable methane. Our results in Phase II shows that there is ample scope for the development of such a process. Low cost Pretreatment and Post treatment options for composite wastewater: It is established in the literature that some non-specific eukaryotic enzymes are able to degrade the difficult to degrade substrates to some extent. A high rate and low cost pre-treatment unit has the potential to increase the performance of the anaerobic reactor to a large extent in a CETP. Routine Monitoring of Microbial Ecology of Bioreactors: In all the biological reactors operated in different countries during Phase II, one critical issue for reactor maintenance and detection of impending reactor failure was ability to monitor the microbial ecology and intervene in a timely fashion. Development of a fast, reliable and simple to use technology in this direction will go a long way in maintaining biological reactors in the developing countries. Our efforts in this direction have yielded encouraging results during Phase II. The method still needs to be tested widely and compared with the genetic tools.

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IITB Report: Characterization of Biosolids fromUASB/Activated Sludge Process/Fluidized Bed Process

Contact Person: C. S. Harendranath Regional Sophisticated Instrument Center Indian Institute of Technology Mumbai, Maharashtra, Pin. 400076, INDIA e-mail: cshnath@iitb.ac.in IIT Bombay participated in ARRPET II in two ways: 1. Collaboration with IITK in the development of UASB for Domestic wastewater treatment. 2. Application of Advanced Imaging and Image Analysis techniques to other biosolids from other NRIs 1. Technology Developed through Research The Technology developed, in collaboration with IITK, was the UASB process as a treatment option for low strength Domestic wastewater. The difficulties usually associated in achieving granulation while faced with low strength wastewaters have been successfully overcome and the UASB, with appropriate additives, has been demonstrated to successfully treat Domestic wastewater. The novelty of the Technology is the use of chitosan and naturally occurring reetha seed extracts as additives while treating low strength Domestic wastewater. The reactors with chitosan and anionic fraction of reetha seed extracts have shown superior granulation and reactor performance. The specific research component involved assessment and evaluation, using advanced imaging [Transmission Electron Microscopy (TEM), Environmental Scanning Electron Microscopy (ESEM) and Electron probe microanalysis(EPMA)] and Image analysis techniques, of the process of granulation (Granule Generation/degeneration/regeneration) in UASB reactor treating low strength Domestic wastewater, and its susceptibility to shock loads. The Key objectives are the establishment of granular sludge with low strength Domestic wastewater, under fluctuating OLR; use of additives for enhancement of granulation and associated reactor performance; and the promotion of granular stability. Electron microscopic (TEM, ESEM, EPMA) and Image analysis protocols were established and periodically corrected taking into account sludge/biosolids variability from time to time. Granulation, a key element in UASB operation/performance, was continuously monitored, assessed and evaluated throughout the 3 year study period and correlated with reactor operation and performance. The reactors operation and performance are divided in to four phases -- Start up/steady state and the other three phases correspond to the periods between the successive introduction of additives. Phase I was characterized by growth of residual aggregates to microaggregates; A shift in microbial population from Methanosaeta to Methanosarcina and back to Methanosaeta ;
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corresponding changes in mean aggregates size, sludge wash out, volatile fatty acid concentration and COD variations. However the reactors achieved steady state and stability during this period. Averaged OLR over the steady state period was ~1.2 KgCOD/m3/d and HRT was 4.8h. During phase II, all the reactors performed similarly and agglomerate sizes larger than 0.3 mm and 0.5 mm started growing. Corresponding COD removal efficiency of about 7080% was achieved in all the reactors in spite of wide fluctuations in influent COD. During Phase III and IV, reactors with chitosan and anionic fractions of reetha extract showed significant enhancement in granulation (Macroaggregates/granules) and corresponding improvements in COD removal efficiency, ECP concentration and Methanogenic activity. Cost Benefit Analysis of the technology developed was carried out, in collaboration with IITK, using activated sludge as the bench mark. Complete list of Research publications : Tiwari M K, Guha S, Harendranath CS, Tripathi S. (2006) Influence of extrinsic factors on granulation in UASB reactor. Appl Microbiol Biotechnol. 71:145-54. Tiwari, M. K.; Guha, S.; Harendranath, C. S.; Tripathi, S. (2005) Enhanced granulation by Natural Ionic Polymer Additives in UASB reactor treating lowstrength wastewater, Water Research, 39: 3801-3810. Tiwari, M. K.; Guha, S.; Harendranath, C. S. (2004) Enhanced granulation in UASB reactor treating low-strength wastewater by natural polymers, Water Science and Technology, 50: 235-240. Singh, R., Quaff, A. R., Guha, S., Harendranath, C. S. Stability of Granules in UASB Reactors Treating Low Strength Wastewater: Flow Variability and Low COD, International Conference on Civil Engineering in New Millennium, CENEM 2007, Jan 11-14, 2007, Bengal Engineering and Science University, Shibpur, Howrah, West Bengal, India. Quaff, A. R., Arokia, V. J., Guha, S., Harendranath, C. S. Population Shift During Startup of UASB Reactor treating Domestic Wastewater, International Conference on Civil Engineering in New Millennium, CENEM 2007, Jan 11-14, 2007, Bengal Engineering and Science University, Shibpur, Howrah, West Bengal, India. Quaff, A. R., Arokia, V. J., Singh, R., Guha, S., Harendranath, C. S. Startup of UASB reactor using low strength domestic wastewater, International Workshop on Biotechnology of Anaerobic Bacteria & Archaea, March 3 & 4, 2006, Agharkar Research Institute, Pune, India. Arokia, V. J., Quaff, A. R., Singh, R., Harendranath, C. S., Guha, S. Application of Image Analysis and Electron Optical Techniques in the Evaluation of Anaerobic Process, International Workshop on Biotechnology of Anaerobic Bacteria & Archaea, March 3 & 4, 2006, Agharkar Research Institute, Pune, India. Singh, A, Shetye, B, Guha, S., Harendranath, C. S. Application of ESEM to study granulation in an UASB reactor treating wastewater, XXVI Annual Conference on Electron Microscopy and Allied Fields, organized by Electron Microscope Society of India at CPRI, Shimla during April 16-18th, 2003 (Best poster Award).

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UASB reactors treating low strength wastewater

Physiochemical Parameters and Bioprocess optimization IITK

Biosolids Characterization - IITB

Sludge samples Regular monitoring

IAS

ESEM and EPMA

TEM

Early stage of granulation

Microaggregate formation Mutual auto attachments of microbes

Ecological examination of granules

Organization and morphological changes in microbes

Spatial distribution of microbes

Image Capturing, optimization, segmentation, processing, Measurement

Presence of inert organic and inorganic particles in the sludge

Size distribution Measurement Fraction of larger agglomerates (macro aggregates).

Comparative analysis of elemental contents

Fraction of smaller agglomerates (micro aggregates) Statistical Analysis

Community composition Microbial Dynamics Competition between Metahnosaeata and Methanosarcina

Arithmetic Mean Geometric mean Coefficiency of uniformity Particles greater than 300 um Particles greater than 500 um

Structure of granules

Early stage of granule degeneration

Microbial Composition Porosity and Gas passage Particle size distribution Surface Morphology

Ruptured cell wall, Cell debris and morphological changes like clumps to single cells - granule disintegration

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2. Pilot scale application

A 100 cu.m./day UASB pilot plant was designed, fabricated and commissioned at Vapi, state of Gujarat, India, in June 2006, in collaboration with IITK and VAPI Waste and Effluent Management Co. Ltd (VWEMCL), and is in operation now. The location where the pilot plant has been commissioned is indicated in the adjoining map. The CETP at Vapi receives effluents from paper pulp, ink, chemical precursors, pesticide manufacturing, fertilizer manufacturing and dye manufacturing industries. The pilot plant was started with above mixed wastewater in addition to the sewage wastewater.

O.1 MLD UASB

Total Height -7.850m Internal Diameter -3.175m Water depth - 6.850m Sludge bed depth- 2.5m

Sludge, Influent and effluent sample was Continuously monitored and assessed

Granule characterization

Physicochemical Analysis

IITB IAS ESEM EPMA TEM DGGE

IITK Methanogenic Activity TSS/ECP/Protein-SugarLipid

Vapi Reactor Operation & Maintainence Physiochemical analysis

Being a joint effort, the roles of IITB, IITK and Vapi are also indicated in the flow diagram. Regular visits to the pilot plant was coordinated by IITB for assessing the reactor performance. A continuous interaction was also maintained with the plant and laboratory personnel at Vapi, and IITK, through regular exchanges of email / courier mail and telephonic conversations. At every 20 days interval, sludge from the reactors was continuously monitored and characterized for its aggregation/granulation, surface morphology, chemistry, community composition and ecological associations. The plant was started with unaltered mixed wastewater from the industrial town of VAPI. The reactor was seeded with a mixture of 60% sludge drying bed and 40% of activated sludge. After the first dose of chitosan addition, there was sudden increase in the arithmetic mean size and the percentage of granules greater than 300 and 500

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um. Targeted Flow rate (4170L/hr) was achieved in about 40 days and the OLR was ~ 2.5 Kg-COD/d/m3. Stepped up flow rate from 800 4170 L/h in the reactor showed a gradual decrease in the mean aggregate size which indicated a need for flow rate reduction. Soon after the flow rate reduction to 1000 L/h, a gradual increase in the mean granule size was noticed until 136 days. Sudden/drastic drop in percentage of granules greater than 300 and 100 um prompted addition of second dose of chitosan. A steady COD removal efficiency of 20 % (corresponding BOD removal efficiency of 30- 35%), attainment of steady and appropriate VFA and pH levels, presence of actively dividing biomass, increased methanogenic activity indicated that the plant was stabilized within three months of reactor operation. TEM studies revealed the presence of various morphotypes reflecting the complex substrate involved and the presence of Methanogens in the sludge samples. Methanogenic Activity and Methane gas production have been noticed. TEM studies also specified that the sludge aggregates in the pilot plant was healthy. Although the UASB reactor seems to be performing better than the existing treatment system at the CETP, Vapi, in view of some of the difficulties faced with the UASB operation/performance, a switch over to Domestic wastewater instead of the complex wastewater has been planned and steps are being examined to enhance the pilot plant performance. This is further addressed in the open issues section.

3. Networking
IITB initiated and carried out networking with the following NRIs within the WWTM and IHWTM groups: 1. 2. 3. 4. 5. 6. 7. Center for Environmental Technology and Management - Vietnam De La Salle University -Philippines University of Morotua - Srilanka Asian Institute of Technology- Bangkok Centre of Paper and Pulp Research Institute Indonesia Indian Institute of Technology Kanpur- India Agarkhar Research Institute

The focus of networking activity was related to extension of application of TEM, ESEM, EPMA and Advanced Image Analysis techniques to other biosolids.
Networking with UoM:

The problem was to evaluate the performance of up flow anaerobic attached growth reactor to remove Fe and Mn from synthetic textile effluents. TEM study helped in the selection of seed sludge for reactor start up. Our studies confirmed that the metal removal, particularly Fe, was mainly due to dismillatory sulfate reduction carried out by SRB. The proliferation of SRB is due to generation of H2S in the reactor. The results of these studies corroborated well with the performance of maximum sulfate and COD reduction. Networking pertaining to wetland studies is in progress.

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Networking with AIT:

The objective of the experiments was to investigate effect of nitrite nitrogen inhibition on the Anammox granule as well as microbial population. IAS and TEM studies, demonstrating effect of nitrite nitrogen concentration on anammox floc /aggregation and presence/ absence of anammoxosome, correlated well with FISH and DGGE results. Further work in this area is in progress.
Networking With DLSU :

The main aim of this work was to select suitable biofilm with appropriate subtrate for Chlordane degradation.
Networking with CENTEMA:

The essential features of the networking was to characterize and evaluate granulation in UASB and SRB reactors treating different wastewater
Networking with CPP:

The main aim of this work was to asses granulation process in UASB reactors treating AOX.
Networking with IITK and UoM-----Characterization of Phytochelatins

IITB was a member of networking group on Characterization of Phytochelatins. The specific role of IITB was to locate and quantify the cadmium concentration in water hyacinth. ICP and EPMA studies confirmed and demonstrated the presence of cadmium in root hair and root tip. The concentration in the root hair is comparatively higher in root hair than in root tip. This was correlated with IITKs result.
Networking with ARI TEM investigations on sludge samples from AOX treating reactor was carried out Networking outside ARRPET

Pilot plant activity was carried out and is in progress with the VAPI Waste and Effluent Management Co. Ltd (VWEMCL). The pilot plant activity has been partially funded by VWEMCL, VAPI in particular the fabrication, installation and day to day operations. In addition networking was carried out with various industries like Maharashtra oil Extractions pvt limited; Reliance dyeing works; Chemtech processors, Soneji food industries, Mumbai; Rajashree sugars and chemical industries, Tamilnadu; Bangalore Dairy; Parle products private limited, Mumbai and TNPCB, Tamilnadu with respect to the survey on agrobased industry effluent treatment plant. Networking with Indira Gandhi centre for Atomic Research (IGCAR) and Madurai Kamaraj University to explore application of confocal laser scanning microscopy and Atomic force microscopy.

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Networking also involved interaction with Central pollution control board, New Delhi, India for EIA activity. Cost Benefit Analysis, which was carried out in collaboration with IITK, involved networking with outside agencies.

4. Dissemination A 3 day workshop on Advanced Analytical techniques was held at IITB during 2-4 April 07. A number of research students and NRIs from IHWTM and WWTM groups participated in the workshop. For further details please visit http://www.arrpet.ait.ac.th. A 5 day Workshop / Training programme was conducted at IITBfor DLSU, UOM and ARI. NRIs in the month of March 2005. Training on IAS, ESEM and TEM techniques was the Highlight of the programme. One week training programme was arranged for Ms Rini Soteppo from CPP on IAS, ESEM, EPMA and TEM techniques in the month of November 2005. A three day workshop on Image analysis and Electron optic techniques was organized for a research student from IITK, during 9-11 September 2006. E-Newsletter for WWTM and HWTM groups was initiated by IITB and issue no 1, August 2005, was designed and posted in ARRPET website. http://www.arrpet.ait.ac.th/wwtm/newsletter1/index.html 5. Capacity Building The following research personnel were trained in various advanced imaging and image analysis techniques and were associated with the project. Bharati Patro, Bhooma Venkatraman, Dr. Aditi Deshpande, Vibha Jayaraj, Shivaji Bhosle, Dillip Kaneskar, Ramesh Mahadik, Neerja Jani and Ajay Singh. A Leica stereo microscope was interfaced with existing LEICA light optical microscope to enable aggregate size distribution analysis of larger granules; A LEITZ PL 2X objective lens was incorporated to extend magnification limits of the light optical microscope; The image analysis software was upgraded time and again to lead to the present AnalySYS 5.0 system; The leica ultramicrotome was upgraded to enable sectioning for TEM experiments; Waterbath, Rotor, pH electrodes, Mini cooler and Micropipettes etc were added to enable initiation of FISH experiments; Various aids like carbon coated grids, tungsten filaments were added to enable EM experiments. 6. Open Issues A major challenge that has remained unresolved in phase II pertains to the successful application of the Technology developed in the laboratory, namely UASB application to Domestic waste water, to Pilot plant studies.

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Some of the issues that need to be addressed are: 1. COD removal efficiency has been limited to about 20%; Possibilty of SRBs inhibiting methanogenic population needs to be explored. 2. Fermentation studies need to be carried out for the influent in order to gain understanding of factors inhibiting granulation. 3. Pretreatment options are to be considered to enable removal of inhibitory components/constituents like Endosulfan, Phenols, oil, greese etc. 4. Experiments to enhance granulation by use of Reetha seed extracts to overcome surfactant and foaming problems. 5. Process optimization remains unresolved in scale up operations. 6. More effective and indepth collaborative work needs to be pursued while applying the Advanced Imaging and Image analysis techniques to other biosolids through networking with other NRIs.

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KMUTT Report: Bioremediation and Reuse of Marine Shrimp Farm Effluent

Contact Person(s): Dr. Piyabutr Wanichpongpan, Dr. Wiwat Ruenglertpanyakul Department of Chemical Engineering Faculty of Engineering King Mongkuts University of Technology Thonburi 126 Pracha-U-thid Road, Tungkru, Bangkok 10140, Thailand e-mail: piyabutr.wan@kmutt.ac.th 1. Technology Developed through Research: Intensive culture of marine shrimp widely practiced in Thailand requires inputs of high-protein diets, with high feeding rates. Most of feeds eaten by shrimps are eventually excreted as metabolic wastes that add inorganic nutrients and organic matter to the water and sediment in the shrimp ponds. The traditional and easiest way for maintaining a good pond water quality is frequent water exchange, but this method extremely pollutes natural water resources and the environment. This project attempted to develop cost-effective methods of improving the shrimppond effluent quality, especially ammonia nitrogen, prior to discharge to the environment with minimum requirement for treatment area. This could be accomplished by utilizing photosynthesis activity of micro-algae developed in shrimp-pond. The new approach has several advantages: (1) relying on the principles of natural ecosystems and therefore was environmental-friendly process; (2) causing no secondary pollution since the produced biomass could be used as fish feed or fertilizer, and (3) enabling the efficient recycling of nutrients contained in the secondary effluents, since the micro-algae are highly effective not only at using the inorganic nitrogen and phosphorus for growth but also at purifying the waste by producing oxygen and removing heavy metals. The treatment of pond effluents was considered as the major issue in this study, while the treatment of the sediment was considered as the minor issue. 1.1 Major Issue: Treatment of pond effluents The study aims to determination of light/dark period for optimum nutrient removal of green algae (Chlorella sp.), find optimum operating conditions of photobioreactor for treatment of effluent in shrimp farming and test the performance of bioremediation in reuse of shrimp-farming effluents. At the first stage, the study was performed at laboratory scale. The green algae, Chlorella sp., was cultivated in Conway medium in 250-mL conical flasks in culture medium with (NH4)2SO4 as N-source and incubated at 30C with shaking speed of 100 rpm. The artificial light (40W fluorescent lamp) supplied 1,100 Lux. The specific ammonia uptake rate (SUR, ammonia reduction rate per micro-algae concentration) was used as index of ammonia treatment efficiency by Chlorella.
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In the larger scale experiments, the effect of lighting period and L/D volume ratio on ammonia removal rate was studied in a 40-L photobioreactor, which consisted of light-receiving area and non-lighting or dark area. A 500-W halogen lamp was used as artificial light-source, which provides at least 1,000-Lux intensity of light over the lighting area. The lighting period was the hydraulic retention time in the lighting area, controlled by the overflow rate of shrimp-pond effluents. The L/D volume ratio was controlled by adjusting the ratio of light-receiving area to dark area. In 200-L experiments, two 200-L plastic ponds interconnected with an overflow pipe were used. One pond is equipped with lighting system, imitating the light exposed area while another was kept dark as no-lighting area. Similar effects of L/D volume ratio on SUR were found in both scale, 40-L photobioreactor and 200L-ponds. The maximum SUR was found at continuous lighting condition and gradually decreased together with decreasing L/D volume ratio. However, photobioreactor required less light receiving volume than the 200-L pond, i. e., only 1:10, at 30 second lighting period to achieve 0.4-0.6 mg-N/mg-CHL.a/h, while 200L pond needed twice higher L/D ratio. These might come from the thin-flat shape of lighting-area and well-mixed condition in the photobioreactor which provided a good light-utilization condition for photosynthesis of the algae. Poorly mixed condition in 200L ponds might cause shading effect and led to reduction in photosynthesis activity. 1.2 Minor Issue: Treatment of pond sediment Sludge or bottom sediment in shrimp ponds after harvesting of shrimp production is composed of polluted organic matter. It stated that each kilogram of pond bottom sludge was composed of; hydrogen sulfide 13.6 mg; ammonia-nitrogen 45.9 mg; orthophosphate 1.2 mg; and organic matter 16.0%. Thus, water extracted from shrimp ponds has the potential to be used as an effective medium for the culture of microalgae. The nutrient-removed sludge can be disposed via land application. Consequently, it can reduce an impact on coastal environment such plankton blooms and eutrophication from the sludge drainage. The three approaches were investigated for sediment treatment, namely, biogas production, nutrient leaching and drying process. The first approach was found to be infeasible. The biogas production ability test using sediment from local shrimp pond showed that the native microorganisms could not produce sufficient biogas either with or without glucose addition (0.1 and 0.5%). Adding biogas producing microorganism into the sediment could also not generate sufficient amount of biogas. The salinity level in sediment might not be suitable for biogas production. In the second approach, nutrient in sediment could be extracted using water. However, the nutrient release rate in this study is very slow, only 0.06 mg-N L-1 d-1 for TAN and 0.03 mg-P L-1 d-1 for phosphate. In the last approach, the population dynamics of microbial community during the sediment drying process had been investigated. The DNA characteristics were used to identify the microorganism species by PCR-RFLP and PCR-DGGE methods. The results indicated that the natural sunlight affected the abundance and microbial community structure in the sediment samples. Direct exposure of sunlight for 3 weeks in summer day (April, 2006) would be sufficient for pond drying process

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to eliminate the undesirable microorganisms. Since UV light cannot penetrate through opaque materials, possible resulting in an inefficient suppression of UVsensitive bacterial species in the bottom zone. Therefore, tilling of bottom sediment during pond drying will enhance efficiency of the exposure. 2. Pilot Scale Application: The aim of this study is to investigate the effect of scale-up on the ammonia removal at larger scale as well as to perform economical analysis of the study at pilot scale. The experiments were performed in a 10-m3 concrete pond (3.54.0 m2 with 0.7m depth) (Fig.1). Airlift pumps were applied around the pond to enhance water circulation and mixing in the pond. Five 500W tungsten lamps were hung over the water surface to supply lighting for the photosynthesis (Fig.1-Left). Each lamp provides minimum 1,000 Lux of light intensity, covering 1.2 m2 of the lighting area above the water surface. Total ammonia nitrogen level was adjusted to 3 mg-N/L by supplement of (NH4)2SO4 at the beginning of prior to the experiment.

Fig.1 The pilot scale application in a 10-m3 concrete pond The results in this experiment showed the same trend as those from the previous study, i.e. at 200-L pond and 40-L photobioreactor. At the optimum L/D ratio, 1/2.3, the maximum specific uptake rate of 0.06 mg-N/mg-Chl.a/h was obtained. Notably, all the specific uptake rates obtained from this experiment were an order of magnitude lower than those obtained from the 200-L photobioreactor study (0.71-0.98 mg-N/mgChl.a/h). Higher depth of pond might produce higher shading effects to the algae. The very high temperatures at the water surface (40-45C) due to the heat from tungsten lights could also negatively affect the ammonia removal by the algae. Additional to lower removal rate, the higher heat loss during lighting made this experiment to be inefficient. To solve a problem of high water temperature and to improve the specific ammonia uptake rate, the 500-W tungsten lamp was replaced by twenty of 16-W fluorescent lamps. The new lamps produce less heat without compromising the lighting efficiency and hence were much more energy efficient. These fluorescent lamps were sealed in glass tubes and immersed under the water surface (Fig.1-Right), to reduce the shading effect near the pond bottom. The results showed that the application of fluorescent lamps could improve the ammonia removal rate. The ammonia uptake rates obtained from fluorescent lamp were about 2.5 to 2.75 time higher than those from 500W lamp 42 ARRPET Phase II

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at the same L/D ratio, with less power consumption. The maximum ammonia uptake rate obtained in this study was 0.15 mg-N/mg-Chl.a/h, which was comparable to the range of ammonia removal rate in laboratory-scaled experiment by many species of cyanobacteria (Spirulina sp., Phormidium sp. and Pseudanabaena sp.), i.e. 0.14 to 0.38 mg-N/mg-Chl.a/h. However, the maximum ammonia removal rate in daytime was as high as 1.8 mg-N/mg-Chl.a/h, which was more 10 times higher than the values obtained under these artificial lights. In summary, the optimum L/D ratio for ammonia removal obtained from this experiment in a 10-m3 pond was 1:3 and excess lighting not only lowers the uptake rate, but also reduces in the NH3 removal efficiency by heat radiation. The application of fluorescent lamps to reduce this radiation can enhance the maximum ammonia removal rate from 0.06 to 0.15 mg-N/mg-Chl.a /h. This is very low compared to that obtained in the laboratory-scale, i.e. 0.71-0.98 mg-N/mg-Chl.a/h. The reason might be the poor mixing in the experimental pond leading to shading effect. Increasing the turbulence of water flow can also enhance this ammonia removal rate to 0.50 mgN/mg-Chl.a/h. 3. Networking: The networking activity with other NRIs and the networking activity with other agencies were listed as follows; 3.1 Within ARRPET 1. Training Workshop on Molecular Techniques, 21-25 March 2005, Asian Institute of Technology, Bangkok, Thailand. 2. International conference on Environmental Management of Urban Infrastructure & Industry in Asian, 11-12 November 2005, CENTEMA-Wageningen UniversityVan Lang University, Ho Chi Minh City, Vietnam. 3. ARRPET Training Workshop on AOX Analysis, 12-14 December 2006, Waste Management Research Centre Faculty of Engineering University Kebangsaan Malaysia, Malaysia. 4. ARRPET II - DLSU 3rd National workshop on Transcending Research Forging Partnership for POPs Management Technology Development, 23-28 January 2005, DLSU, Manila, Philippines. 3.2 Outside ARRPET 1. Chaipattana Foundation: The Foundation agrees to support our research group and KMUTT to develop research facilities for pilot scale test and demonstration farm in Bang Khun Tien, Bangkok, Thailand. 4. Dissemination: There were 2 national workshops held and 4 technical reports published during this project period. 4.1 Workshops 1. The national workshop in the title of Sustainable Shrimp Farming was held on June 27-28th, 2005 at The Rock Garden Beach Hotel, Rayong, Thailand.

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2. The national workshop in the title of Sustainable Aquaculture Engineering Approach was held on February 8-9th, 2007 at Hin Nam Sai Suay Hotel, Hua Hin, Thailand (cooperated with KMUTT&CENTEMA). 4.2 Publications 1. Attasart S., Ruenglertpanyakul W., Wanichpongpan P. (2005) Utilization of Chitosan for Organic Shrimp Production, J. of Metals, Materials and Minerals, Vol.15, No.1. 2. Attasart S., Ruenglertpanyakul W., Wanichpongpan P. (2005) Bioremidiation and reuse of marine shrimp farm effluent, International conference on Environmental Management of Urban Infrastructure & Industry in Asian, 11-12 November, Ho Chi Minh City, Vietnam. 3. Wiwat, R., Piyabutr, W. (2005) Bioremediation of shrimp farming effluents by using microalgae, The 2nd national conference on algae and plankton, ChiangMai University, 23-25 March, Holliday Inn Hotel, Chiang-Mai, Thailand. 4. Boonapatcharoen N., Wanichpongpan P., Ruenglertpanyakul W., Techkarnjanaruk S. (2006) Effect of Natural Sunlight on Microbial Population in Shrimp Farming Sediment on The International Conference on Environment and Public Health Management: Aquaculture and Environment, 6-12 December, Hong Kong. 5. Capacity Building: 5.1 Manpower There are 5 research staffs and 1 Ph.D. student in this program. 1. Asst. Prof. Dr. Piyabutr Wanichpongpan (Ph.D. Eng.) 2. Asst. Prof.Dr. Wiwat Ruenglertpanyakul (Ph.D. Technische Chemie) 3. Dr. Somkiet Techkarnjanaruk (Ph.D. Molecular Microbiology) 4. Dr. Wattanee Sriwatanapongse (Ph.D. Eng) 5. Ms. Onamon Laopitinan (M. Eng.) 6. Mr. Shewin Attasat (Ph.D. Student) 5.2 Laboratory Infrastructure The lists of minor equipments purchased in this period are as followed: 1. Table top centrifuge (Gemmy Industrial Corp., model: PCL-012, Taiwan) 2. Automatic autoclave (Rexall Industies Co. Ltd., model: LS-2D, Taiwan) 3. Portable personal computer (Fufitsu Lifebook S211) 6. Open Issues: Though the algae showed high potential in nitrogen removal, the excess produced algae needs to be harvested. Using filters or sedimentation to remove the algae was investigated but it was found to be infeasible due to extra required area or equipment to harvest low value-added algae. Fish or bivalves could be culture together with shrimp, but there might be some problem of competition on shrimp feed. Another alternative is to develop a productive wetland for treating this excess algae as well as other left-over nutrient. The productive wetland does not only remove nutrients but could use to produce high value-added aqua-product as well. A good proportion of production area for each species could maximize the total production as well as the profit with minimum environmental impact.

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