Specialist Group on ANAEROBIC DIGESTION Newsletter - IWA

<strong>Specialist</strong> <strong>Group</strong> on 

ANAEROBIC DIGESTION 

Newsletter 

June 2009 

CONTENTS 

Chairman’s note 2 

Contributions to the Newsletter 2 

Management committee of the AD <strong>Specialist</strong> <strong>Group</strong> 3 

How to join the <strong>Specialist</strong> <strong>Group</strong> on Anaerobic Digestion 3 

Update from the Task <strong>Group</strong>s on Anaerobic Digestion 4 

News from the world on Anaerobic Digestion 5 

AD people on the move 6 

Past and coming conferences and events 8 

Books and publications 11 

PhD theses on anaerobic digestion 13 

Ongoing anaerobic digestion projects 29 

Anaerobic URL sites 33 

Disclaimer: This is not a journal, but a Newsletter issued by the IWA <strong>Specialist</strong> <strong>Group</strong> on Anaerobic Digestion. 

Statements made in this Newsletter do not necessarily represent the views of the <strong>Specialist</strong> <strong>Group</strong> or those of the 

IWA. The use of information supplied in the Newsletter is at the sole risk of the user, as the <strong>Specialist</strong> <strong>Group</strong> 

and the IWA do not accept any responsibility or liability. 

IWA <strong>Specialist</strong> <strong>Group</strong> on Anaerobic Digestion 

June 2009 Newsletter 1

CHAIRMAN’S NOTE 

Welcome to the 2009 Anaerobic Digestion <strong>Specialist</strong> <strong>Group</strong> Newsletter, and a 

new specialist group management committee. As most of you probably know, 

the AD specialist group is one of the largest in IWA, with over 1000 members, 

and has a massive impact on public policy, and the sustainable application of 

wastewater around the world. 

The outgoing committee, headed by Jules van Lier, and Sergey Kalyuzhnyi 

were a key part of the success and activity of the specialist group, and we now 

have some big shoes to fill. Gladly, Jules has agreed to continue his input to the 

specialist group as Western Europe representative. Jules is one of four original 

management committee members who continued, out of 20 members in total. It 

is very exciting that we have so many new, and early career specialists in the 

management committee, to ensure inspired leadership for anaerobic digestion for 

the future. 

We also have a lot of exciting events coming up in the near future. AD12, the 

major anaerobic conference will be held in Guadalajara, Mexico in late October 2010. In past congresses, we 

have seen the emergence of major themes, including hydrogen production in Montreal at AD10, and more 

recently, bio-electrochemical systems at AD11 in Brisbane. Supporting this, we have AD sponsored specialist 

conferences, including WWTmod2010, in March 2010 in Canada, an area for emerging modelling topics, and 

Anaerobic Digestion of Solid Wastes and Energy Crops, to be held in 2011, in Vienna. Along with these 

<strong>Specialist</strong> <strong>Group</strong> conferences, there will be a number of new initiatives directly from IWA, aimed at increasing 

collaboration between specialist groups. Together with the rest of the management committee, and IWA as a 

whole, we thank you, our specialist group members for your support through the election process. I look 

forward to meeting old and new friends in the specialist group very soon, but in the meantime, please enjoy our 

latest newsletter. 

CONTRIBUTIONS TO THE NEWSLETTER 

Contributions to the Newsletter should be addressed to the Newsletter Editor or alternatively to the <strong>Group</strong> 

Chairman or to the <strong>Group</strong> Secretary. Please submit your contributions by e-mail and in Microsoft Word format. 

<strong>Group</strong> Chairman <strong>Group</strong> Secretary Newsletter Editor 

Dr Damien Batstone 

Advanced Water Management Centre 

(AWMC) 

The University of Queensland 

St Lucia, 4072 

Australia 

fax: +61-7-33654726 

e-mail: damienb@awmc.uq.edu.au 

Dr Henri Spanjers 

Lettinga Associates Foundation 

(LeAF) 

P.O. Box 500 

NL-6700 AM Wageningen 

The Netherlands 

Tel: +31 317483202/+31 317482023 

e-mail: henri.spanjers@wur.nl 

Dr Jorge Rodríguez-Rodríguez 

Sustainable Environment Research Centre 

(SERC) 

University of Glamorgan 

Pontypridd CF37 1DL 

United Kingdom 

Tel: +44(0)1443 65 4282 

email: jrodrigu@glam.ac.uk 

Editor’s note: The best care was taken to avoid typing errors during the editing of the contributions received 

from SG members. However should the reader find any mistake please report it to the editor and it will be 

corrected in the next issue. 


2 June 2009 Newsletter

MANAGEMENT COMITEE OF THE AD SPECIALIST GROUP 

A renovated Management Committee for the IWA Anaerobic Digestion <strong>Specialist</strong> <strong>Group</strong> has been elected early 

this year. The composition of the new committee is as follows: 

MANAG. COMMITTEE ROLE PERSON ELECTED E-MAIL 

Chair Damien Batstone damienb@awmc.uq.edu.au 

Secretary Henri Spanjers henri.spanjers@wur.nl 

Newsletter Editor Jorge Rodríguez-Rodríguez jrodrigu@glam.ac.uk 

REGIONAL REPRESENTATIVES 

Northern Europe Irini Angelidaki ria@er.dtu.dk 

Western Europe 

Jules B. van Lier 

Alan Guwy 

j.b.vanlier@tudelft.nl 

ajguwy@glam.ac.uk 

Southern Europe 

Joan Mata-Álvarez 

Jean-Philippe Steyer 

jmata@ub.edu 

steyer@ensam.inra.fr 

Central & Eastern Europe Pavel Jenicek 

Eberhard F. Morgenroth 

pavel.jenicek@vscht.cz 

eberhard.morgenroth@eawag.ch 

North America 

David Bagley 

Viviane Yargeau 

bagley@uwyo.edu 

viviane.yargeau@mcgill.ca 

Central America Germán Buitrón gbuitronm@ii.unam.mx 

South America Carlos Chernicharo calemos@desa.ufmg.br 

Middle East Moktar Hamdi moktar.hamdi@insat.rnu.tn 

Central Asia Makarand M. Ghangrekar ghangrekar@civil.iitkgp.in 

Asia Raman Saravanane rsaravanane@pec.edu 

Asian Islands Hiroshi Tsuno hs-tsuno@water.mbox.media.kyoto-u.ac.jp 

Australia Jurg Keller j.keller@uq.edu.au 

HOW TO JOIN THE SPECIALIST GROUP ON ANAEROBIC DIGESTION 

All members of IWA are welcome to join the <strong>Specialist</strong> <strong>Group</strong>. With the IWA registration, you can indicate 

which <strong>Specialist</strong> <strong>Group</strong>(s) you wish to join. 

If you wish to become member of the <strong>Specialist</strong> <strong>Group</strong> on Anaerobic Digestion and you did not have indicated it 

on your IWA registration form, please contact the Chairman or the Secretary. 

You may also directly contact IWA: 

Keith Robertson (keith.robertson@iwahq.org.uk) or Frances Lucraft (frances.lucraft@iwahq.org.uk) 



UPDATE FROM THE TASK GROUPS ON ANAEROBIC DIGESTION 

IWA Task <strong>Group</strong> on Mathematical Modelling of Anaerobic Digestion 

The IWA Anaerobic Digestion Modelling Task <strong>Group</strong> is currently not active, but we have observed a number of 

developments in the area. A workshop was held at WWTmod 2008 in Quebec, Canada. This was aimed at 

identifying some of the key limitations of the ADM1 for practical use. The two major issues identified were 

characterisation of influents, and the physicochemical system. In particular, for the latter, it was identified that 

phosphorous still could not be effectively modelled in anaerobic digesters (or in aerobic systems). The IWA 

Benchmarking Model Taskgroup has partly addressed the first issue as Nopens, et al. (2009) "An ASM/ADM 

model interface for dynamic plant-wide simulation" Wat. Res. 43(7): 1913-1923. This provides an input model 

for relatively straight forward definition from activated sludge, and can be readily adapted to other substrates. 

The main task remaining is providing reference applications of this input model to areas other than activated 

sludge. The issue of advanced physicochemical models is more involved, and some of the challenges have been 

further outlined in Batstone, D.J. (2009) “Towards a generalised physicochemical modelling framework” 

Reviews in Environ. Sci. Biotech 8(2):113. 

Damien Batstone, Chair of ADM Task <strong>Group</strong> 

IWA-AD Task <strong>Group</strong> on Harmonization of Anaerobic Biodegradation, Activity and Inhibition Assays 

(TG ABAI) 

The ABAI TG has the objective to harmonise anaerobic biodegradability, activity and inhibition assays. The 

group met last time for a workshop in Prague, where a report was written summarising the discussions during 

the ABAI TG. Based on the work of ABAI TG a paper was presented by at the Solid waste conference by David 

Bolzonella in Hemet, Tunesia. The paper has been recently published: 

I. Angelidaki, M. Alves, D. Bolzonella, L. Borzacconi, J. L. Campos, A. J. Guwy, S. Kalyuzhnyi, P. 

Jenicek and J. B. van Lier (2009). Defining the biomethane potential (BMP) of solid organic wastes and 

energy crops: a proposed protocol for batch assays. Water Sci. Technol. 59(5), pp. 927-934 

Irini Angelidaki, Chair of the ABAI-TG 



NEWS FROM THE WORLD ON ANAEROBIC DIGESTION 

Lee Kuan Yew Water prize 2009 awarded to Gatze Lettinga 

(from LeAF Newsletter March 2009) 

We are happy to announce that Gatze Lettinga is awarded the 

Lee Kuan Yew Water prize 2009. The Lee Kuan Yew Water 

prize is an annual award for “outstanding contributions 

towards solving global water problems by applying 

groundbreaking technologies or implementing innovative 

policies and programmes which benefit mankind”. The prize 

comprises a gold medallion, an award certificate and a cash 

prize of S$300000. The prize is named after former 

Singapore Prime Minister and present Minister Mentor Lee 

Kuan Yew, who has been exceptionally important for 

Singapore in attaining a sustainable water supply. 

Gatze Lettinga receives the award for his “breakthrough, 

environmentally sustainable solution for the treatment of 

used water using anaerobic technology”. The jury recognizes the revolutionary aspects of the treatment concept 

that enables cost-effective (pre-) treatment of wastewater, combined with the production of renewable energy, 

fertilizer and soil conditioner. Furthermore, the jury appreciates the fact that the technology was never patented, 

which makes the technology universally available. As a consequence anaerobic wastewater technology has been 

widely applied for both industrial and municipal wastewater. 

This is the second time that the prize has been awarded. Gatze Lettinga was chosen from a total of 39 

nominations from 19 different countries. He will receive the award on June 24 in the official residence and 

office of Singapore’s president during the Singapore International Water Week (SIWW) 22-26 June 2009. In 

addition, Gatze Lettinga will deliver the Singapore Water Lecture on June 23. 

The SIWW is a global platform for experts in different fields to discuss ongoing issues in the water world. The 

theme in 2009 is “SustainableCities – Infrastructure and technologies for Water”. During this event we will 

organise a special session dedicated to sustainable technology development with Gatze Lettinga as honoured 

guest. 

Gatze Lettinga was professor “Anaerobic Treatment Technology and Reuse” at the subdepartment of 

Environmental Technology of Wageningen University from 1988 to 2001. In 1997 and together with professor 

Jules van Lier, he founded LeAF, of which he was board member till July 2004. Nowadays, he is still active as 

advisor of the board of LeAF (amongst his numerous other activities). Gatze Lettinga’s achievements have been 

honoured by several awards. Only two years ago he received the prestigious Tyler prize for Environmental 

Achievements in Los Angeles, USA. 

For more information on the Lee Kuan Yew Water prize and the Singapore International Water Week: 

http://www.siww.com.sg/ 



AD PEOPLE ON THE MOVE 

OPEN POSITIONS 

21 month postdoctoral position at the Laboratoire de Biotechnologie de l’Environnement (LBE- 

INRA), Narbonne – France on the topic of “Optimisation of biomass pretreatment and 

hydrolysis to maximise biogas production from agricultural wastes” 

The work will be undertaken at the Laboratoire de Biotechnologie de l’Environnement (LBE). The candidate 

will take advantage of working in a multi-disciplinary group with competences in microbial engineering, process 

engineering, modeling, automatics and microbial ecology. 

This post-doc position will be included in a research collaboration with Campinas University (Unicamp, INRA 

FASESP project). 

This project will have three main objectives: 

1- To maximise biogas (biomethane) production from several lignocellulosic residues by the development of 

physico-chemical and/or enzymatic pre-treatments, 

2- To optimise pre-treatments and find relationships between optimal pre-treatment conditions and biomass 

characterisation 

3- To determine bio-physico-chemical indicators of anaerobic biodegradability (or biomethane potential). 

Besides, batch anaerobic digestion runs which will allow us to assess biomethane production from raw and 

pretreated biomass. Experiments will also concern : 

- biomass characterisation according to classical methods used within residues treatment domain. Innovating 

methods or methods taken from other scientific fields are also forecasted. They may include biochemical, 

chemical, physical or spectroscopic methods. 

- biomass pretreatments which may include physical, thermo-chemical and enzymatic techniques, 

- modeling of pre-treatments and anaerobic digestion to describe relations between biomass characterisation 

and optimal pretreatment conditions and methane production. 

Starting date from September 1st 2009 Duration 21 months 

Expected competence of the candidate: 

- Candidates must hold a PhD in process engineering or bioengineering 

- Skills in lignocellulosic biomass treatment will be a plus 

- The ability to communicate and publish in English is mandatory 

- Candidates should combine autonomy and team working abilities. 

Candidature conditions: 

• Doctor at recruitment date (French doctorate, PhD or foreign doctorate of equivalent level) for less than 5 

years, 

• To fill an application on INRA Web site ( www.inra.fr ) before July 31 st . 

Contact: Dr. Hélène Carrère (carrere@supagro.inra.fr) and Dr. Jean-Phi Steyer (steyer@supagro.inra.fr) 

INRA, UR 050, Laboratoire de biotechnologie de l’Environnement, Avenue des Etangs, 11100 Narbonne, 

FRANCE. http://www.montpellier.inra.fr/narbonne 

Open PhD positions in Hamburg University 

Investigation of the microbial diversity in anaerobic digesters for renewable biomass (sugar beet silage) by FISH 

technique. 

Investigation of the microbial diversity in anaerobic digesters for renewable biomass (sugar beet silage) by the 

RFLP technique. 

Contact: Prof. Dr. Paul Scherer 

Hamburg University of Applied Sciences 

Lohbruegger Kirchstr. 65 

21033 Hamburg-Bergedorf (Germany) 

Tel.: +49/40/42875-6355 or -6351 

Fax: +49/40/42875-6359 

www.haw-hamburg.de/4357 (profile) 

Paul.Scherer@haw-hamburg.de 



CHANGES OF AFFILIATION 

Largus (Lars) T. Angenent has joined the Department of Biological and Environmental Engineering at Cornell 

University at the rank of Associate Professor after 6 years on the faculty at Washington University in St. Louis. 

Dr. Angenent holds a Ph.D. in Environmental Engineering from Iowa State University, an M.S. in 

Environmental Technology/Microbiology from Wageningen University (formerly Wageningen Agricultural 

University), Wageningen, The Netherlands, and a B.S. in Environmental Sciences from Wageningen University. 

Dr. Angenent’s research focuses on optimizing biological anaerobic fermentation processes to foster undefined 

mixed cultures that convert organic wastes into bioenergy carriers, such as methane, electrical current, hydrogen, 

or precursors for the liquid biofuel butanol. In this area, his lab focuses on improving the performance and 

stability of anaerobic digesters, on novel microbial fuel cell configurations, and on the optimization of anaerobic 

fermentation or respiration. When necessary, molecular biology techniques are used in conjunction with longterm 

bioreactor studies. Dr. Angenent is a member of the International Water Association, Association of 

Environmental Engineering and Science Professors, American Society for Microbiology, American Association 

for the Advancement of Science, American Institute of BioEngineering. 

Samir K. Khanal recently joined the Dept. of Molecular Biosciences and Bioengineering, University of Hawaii 

at Manoa as an Assistant Professor. Prior to joining University of Hawaii, Dr. Khanal was a Research Assistant 

Professor in the Department of Civil, Construction and Environmental Engineering at Iowa State University, 

Ames, IA. Dr. Khanal’s research focuses on production of biofuels/Bioenergy and valuable bio-based products 

from agri/forest-residues and wastes. Dr. Khanal is a co-recipient of R&D 100 Award, 2008 Grand Prize for 

University Research presented by the American Academy of Environmental Engineers and International Water 

Association (IWA) Project Innovation Award.. Dr. Khanal also recently published a book " Anaerobic 

Biotechnology for Bioenergy Production: Principles and Application (Wiley-Blackwell Publishing). Dr. Khanal 

is a registered engineer in the state of Iowa. 

Titia de Mes has worked at DHV B.V. as a Process Design Engineer - Wastewater Treatment, Amersfoort, 

Netherlands. In August 2008 I took up a position within Atkins, Dubai, UAE as Senior Wastewater Engineer. In 

the Netherlands wastewater effluents are often discharged into streams and not reused. In the Middle East, 

potable water is mainly obtained from energy intensive desalination processes, the reuse of wastewater effluent 

is very feasible. One of my roles within Atkins is to advise our clients on water reuse schemes, appropriate 

treatment and water and energy saving options. titia.demes@atkinsglobal.com 

Eberhard Morgenroth has served on the faculty of the University of Illinois at Urbana-Champaign since 2000. 

Starting in August 2009 he will take over a professor position at ETH (Swiss Federal Institute of Technology 

Zürich) and EAWAG (Swiss Federal Institute of Aquatic Science and Technology). His new contact information 

are: eberhard.morgenroth@eawag.ch 



PAST AND COMING CONFERENCES AND EVENTS 

An experts' colloquium on anaerobic technology: 

Sustainable Energy Production and Wastewater Treatment 

Keynote Speaker: Prof. Dr. Gatze Lettinga 

The Experts’ Colloquium will be held on the morning of Thursday, 25 th June 2009 at the SUNTEC Singapore 

International Convention & Exhibition Centre. Attendance is by invitation only. Kindly contact Ms. Tai Xiang 

(xtai@wspc.com) for inclusion into the invitation list. 

The Nanyang Environment and Water Research Institute (NEWRI) and Lettinga Associates Foundation 

(LeAF), together with World Scientific Publishing, are proud to announce An Experts' Colloquium on 

Anaerobic Technology - Sustainable Energy Production and Wastewater Treatment. This Colloquium, held in 

honor of Prof. Dr. Gatze Lettinga, winner of the 2009 Lee Kuan Yew Water Prize, aims to bring together leaders 

in anaerobic technology to discuss the sustainability and future of Anaerobic Technology. 

Held in conjunction with the Singapore International Water Week 2009, an international panel of leading 

academics and industry speakers will be touching on various topics in anaerobic technology. The goal of the 

workshop is to share with stakeholders a more sustainable approach in environmental engineering, focusing on 

the state of the art, potentials & challenges of implementing modern high-rate anaerobic treatment systems in 

integrated settings for applications in Asia and elsewhere. The Colloquium’s Keynote Speaker, Prof. Dr. Gatze 

Lettinga will share his insights on the topic “The Ever-Improving Digestibility of High Rate Anaerobic 

Wastewater Treatment”. 

The forthcoming journal, Advances in Environmental Research and Development (AERD) will also be 

launched during the colloquium. AERD aims for rapid dissemination of original and high quality technical 

papers which may be in materials, analytical techniques, water and wastewater treatment, water reclamation, 

water quality, modeling of quantity and quality, air pollution management and control, soil remediation, and 

wastes management and resource recovery. 

Annual Anaerobic Treatment Short Course in USA 

The annual short course titled "Anaerobic Treatment of High-Strength Industrial Waste" will be held September 

16-17, 2009 at Marquette University in Milwaukee, Wisconsin, USA. Speakers will include Drs. Willy 

Verstraete (Gent University), Daniel Zitomer (Marquette University) and Dennis Totzke (Applied Technologies, 

Inc.). 

Information will be presented regarding sustainability and greenhouse gas emission avoidance, biogas 

utilization, anaerobic treatment fundamentals, operating strategies, design/construction/start-up guidelines and 

case studies of operating anaerobic treatment systems. 

For more information, see http://www.marquette.edu/ANT after July 15 or email Daniel Zitomer 

(daniel.zitomer@mu.edu) 

III Simposio colombiano sobre Biotecnología Ambiental y III Simposio Colombiano sobre 

Digestión Anaerobia 

Late October, 2009. Auditorio Mario Laserna, Universidad de los Andes 

Organisers: Centro de Investigaciones en Ingeniería Ambiental [CIIA] Universidad de los Andes y Red 

Colombiana de Biotecnología Ambiental [RECBAM] 

More info: http://IIIsimposiobiotecnologia.uniandes.com 



The International Workshop on Strategy for Anaerobic Biotechnology 

24-27 November 2009 Xi'an Nan Yang Hotel, Xi’an, China. Hosted by: Xi’an Jiaotong University, Xi’an, China 

Anaerobic treatment has been accepted as an important technology which has been playing a key role in 

sustainable development since the past thirty years. A symbol for this is that the very prestigious “Tyler Prize of 

Environmental Achievement” (April, 2007) and the “Lee Kuan Yew Prize”(June, 2009) awarded Professor G. 

Lettinga, the main inventor of UASB and the promoter of the other innovative concepts on anaerobic 

technology. To push forward the application of this sustainable technology, an international workshop on 

anaerobic biotechnology will be held on November 24-27, 2009 in Xi'an, China. It will also be good for the 

people who are engaged in this field to meet each other, exchange their experiences and ideas. On the occasion, 

famous experts and scholars will give interesting reports. 

Topics: 

1. The research and application of anaerobic biological in industrial and domestic wastewater treatment. 

2. Key technology and equipment on anaerobic wastewater treatment 

3. The research and application of anaerobic digestion of solid wastes 

4. Strategy for development of anaerobic technology 

5. Conversion and utilization technology of energy from anaerobic biological treatment. 

6. Others 

Key Dates: 

April 20, 2009: First Announcement and call for papers 

July 20, 2009: Deadline for submission of abstracts and the second announcement 

October 8, 2009: Deadline for submission of full papers 

November 1, 2009: Third announcement 

November 23, 2009: Registration at Xi’an Nanyang Hotel 

November 27-28: Departure 

Before 23 or after 27 of November, visit to Tsinghua University in Beijing. 

Fee: (1) No registration fee. (2) Free industrial visit 

International Workshop on Anaerobic Digestion: An old story for today and tomorrow 

10-11 th December 2009, Narbonne, France 

From A. Volta to R. Moletta, what have we learnt And how could this be useful for tomorrow 

Dr. René Moletta, the founder of the “Laboratoire de Biotechnologie de 

l’Environnement” (LBE-INRA - http://www.montpellier.inra.fr/narbonne) has 

been active for more than 30 years in promoting Anaerobic Digestion all over the 

world, both at a scientific level and in an industrial perspective. Before he retires, 

former colleagues wish to thank him in organising a special 2 day workshop on 

Anaerobic Digestion, An Old Story for Today and Tomorrow. This workshop will 

be a unique opportunity to listen to world wide recognised top scientists, to hear 

their great expertise in the field, to share their experience and their view of history, 

and to discuss with them about future in a friendly atmosphere. This workshop is 

also special in the sense that there will be no deadlines for sending an abstract… 

just a place to be and not to miss ! Both from an academic and industrial 

perspective! 

Registration is limited to 150 persons and early registration is 

recommended. It will take place in Narbonne, France, on the 10th and 

11th of December 2009. Narbonne is a lovely and sunny city located 

in the south of France and has a great history since the Roman time. 

The workshop will be help in the “salle des synodes”, a charming hall 

in the old palace of the archbishops, downtown Narbonne. 

For more information and details, please contact: 

Jean-Phi Steyer (steyer@supagro.inra.fr) or Romain Cresson 

(cresson@supagro.inra.fr) 



OLIVEBIOTEQ (Seminar on biotechnology and olive products quality in the Mediterranean) 

The 3rd Edition of OLIVEBIOTEQ, a congress-workshop dedicated to the olive industry, will take place next 

15-19 December 2009 in Sfax, Tunisia. This international congress covers all aspects of this industry from the 

agronomy of olive tree culture to the technology of olive transformation (olive canning and olive oil production) 

and the management of the resulting sub-products (three phase olive mill wastewater and husk, two-phase olive 

mill waste, wastewater from olive canning, biomass from olive tree pruning). Olivebioteq is one of the most 

important events for the actors of this industrial sector in the Mediterranean basin. Beside of being an important 

economic activity for several Mediterranean countries, olive industry is also one of the most polluting in the 

same area. Until now, no satisfactory solution has been applied by this industry for the protection of the 

environment. Direct anaerobic digestion or co-digestion of its sub-products is an alternative that still looks for 

recognition and acceptation by this industry. The 3rd Olivebioteq is then a good opportunity to promote the 

activities of those of you who work in this field and enters perfectly in the frame of the topic number 6: “New 

uses and valorisation alternatives of olive tree products (nutrition, health, cosmetics, energy, environment ...)”. 

More information about the congress is available at the following link: 

http://www.iresa.agrinet.tn/oliveteq/1-en.html 

The call for papers is opened until June 30, 2009. A commercial exhibition will be set up in parallel to the 

workshop-congress. 

WWTmod2010, the 2 nd IWA/WEF Wastewater Treatment Modelling Seminar 

It will be held in Mont-Sainte-Anne, Québec, Canada, March 28-30, 2010. This is one of the principal seminars 

for model development and solution policy, and is supported by the IWA AD specialist group. Further 

information can be found here: 

http://www.wef.org/ConferencesTraining/ConferencesEvents/WWTmod2010/ 

12 th World Congress on Anaerobic Digestion 

As you may know, the next big event of the anaerobic community will be in Guadalajara, Mexico next year. The 

IWA <strong>Specialist</strong> <strong>Group</strong> on Anaerobic Digestion selected that city as the venue for the 12 th World Congress on 

Anaerobic Digestion. 

The organizing committee for AD 12 has been working on site in 

order to define the date, hotel, technical and touristic visits, to name a 

few of the initial decisions to be made. We can inform that the local 

venue will be Fiesta Americana Hotel, very well located in Guadalajara 

and that the congress will be held from Sunday October 31 st to Thursday 

November 4 th 2010. Technical visits will take place on Friday 5 th . 

The institutions involved in the organization are the Institute of 

Engineering of the National University of Mexico (UNAM) the 

University of Guadalajara, the Scientific and Technology Research Institute of San Luis Potosi (IPICyT) and the 

Metropolitan University (UAM). 

Guadalajara is the capital city of Jalisco state. It is the second / third most important city of Mexico. This 

region is the cradle of Tequila and Mariachi, two symbols of Mexico. Some Tequila factories have installed 

anaerobic treatment plants, so the visit to the town of Tequila will be a most attractive technical-tourist visit, just 

60 km from Guadalajara. 

We are expecting around 600 delegates to AD 12, due to the traditional high technical level of the congress, 

but also to the tourist attractions that Jalisco and the whole Mexico offers: variety of cultural expressions such as 

archaeological sites and colonial cities, renowned folk handcrafts, plenty of natural beauties, beaches, 

mountains, deserts, jungle, a diversity of cuisine styles, and the warmth of the welcoming characteristic of its 

people. 

The program and the scientific committee will be defined during this June, as the invitation and confirmation 

process is under way. The call for papers and the web site of the congress will be announced in a few weeks. A 

small commercial exhibition will also take place. We count on your participation. 



BOOKS AND PUBLICATIONS 

Anaerobic Biotechnology and Odor/Corrosion Control for Municipalities and Industries 

Author: R. E. Speece, 

Vanderbilt University (616 pages) Price $135. US. 

Arehae Press, 5840 Robert E. Lee Drive, Nashville, TN USA 37215 

Anaerobic Biotechnology and Odor/Corrosion Control for 

Municipalities and Industries is a unique resource prepared to provide 

on the job training for water quality specialists seeking up-to-date 

answers for optimizing design and operation of municipal sludge 

digesters and industrial anaerobic treatment processes preventing odor 

and corrosion formation in collection and treatment systems 

This book (2008) is an expanded version of Anaerobic 

Biotechnology for Industrial Wastewaters by R. E. Speece (1996), 

containing 170 new pages. 

In Part I as chapters 1-7, and largely focused on municipal 

applications. One chapter explores global issues of CH 4 and H 2 S. There 

are 70 pages on the anaerobic digestion of municipal sludges and a 

chapter on biogas utilization is included. Extensive coverage is included 

of odor/corrosion control in municipal collection systems. There are two 

general approaches to odor control: allowing the hydrogen sulfide to 

form and then attempting to treat it or preventing its generation in the 

first place. A recent technology, Superoxygenation, using the second 

approach is included as Chapter 7 because of its proven effectiveness in 

odor and corrosion prevention as demonstrated in full-scale installations. 

When the generation of hydrogen sulfide is eliminated, so also is the 

biogenic formation of sulfuric acid. Updated content of the earlier text is included in Part II as chapters 8-19, 

pertaining to industrial effluent treatment. 

The author's decades of teaching, research and consulting experience in adapting the remarkable possibilities 

of anaerobic biotechnology to both municipal and industrial wastewater reclamation, eminently qualify him to 

compile comprehensive research and on-site reports currently available, and collate them into this valuable 

reference. Anaerobic processes at present are very useful in protecting and enhancing our environment. Since 

regulatory issues drive technology, newer anaerobic configurations and protocols are becoming widely used 

successfully amidst present day shifts in financial advantage and heightened public outcry against odor and 

corrosion problems. 

Key Topics 

• Preventing odors and corrosion in municipal collection systems 

• Assessing the proper role of microbial consortia proximity 

• Incorporating recuperative thickening to maximize reactor volume usage 

• Optimizing thermophilic digestion to meet EPA standards for Class A biosolids 

• Biodegrading toxicants 

Special Features 

• Text boxes throughout for clarity and factual retention 

• In depth study of methane-producers and hydrogen sulfide-producers 

• Inclusion of Superoxygenation technology option for prevention of odors and corrosion 

To Order: 

Contact Archae Press (www.archaepress.com) 

Archae Press 

5840 R. E. Lee Drive 

Nashville, TN 37215 

615-665-0847 

dick.speece@vanderbilt.edu 



Anaerobic Biotechnology for Bioenergy Production: Principles and Application 

Author: Samir K. Khanal, Ph.D., P.E. (University of Hawai'i at Manoa) 

Publisher: Wiley-Blackwell (www.wiley.com) 

Publication date: October 2008 

ISBN: 978-0-8138-2346-1 

Anaerobic biotechnology is a cost-effective and sustainable means of 

treating waste and wastewaters that couples treatment processes with the 

reclamation of useful by-products and renewable biofuels. This means of 

treating municipal, agricultural, and industrial wastes allows waste products 

to be converted to value-added products such as biofuels, biofertilizers, and 

other chemicals. Anaerobic Biotechnology for Bioenergy Production: 

Principles and Applications provides the reader with basic principles of 

anaerobic processes alongside practical uses of anaerobic biotechnology 

options. This book will be a valuable reference to any professional currently 

considering or working with anaerobic biotechnology options. 

• Coverage of basic Anaerobic Biotechnology principles 

• Practical applications of principles and processes 

• Thorough coverage of cost-effective and sustainable means of treating 

waste water and resource reclamation 

The details about book can be found at: 

http://www.wiley.com/WileyCDA/WileyTitle/productCd-0813823463.html 



PhD THESES ON ANAEROBIC DIGESTION 

Biomass Granulation and Treatment of Sewage using UASB Reactor 

by Puspendu Bhunia 

Institution: 

Civil Engineering Department 

Indian Institute of Technology 

Kharagpur-721 302, WB, India 

Supervisor: 

Dr M.M. Ghangrekar 

Abstract: 

Biomass granulation in upflow anaerobic sludge blanket (UASB) reactor still remains a challenging task while 

treating many wastewaters. This study enlightens the criteria’s for the determination of minimum size of 

granules, required to gain advantage of granular sludge configuration and variation of different properties with 

size of granules. Based on sedimentation theory, this study suggests 0.34 mm as the minimum size of granules 

for sludge with specific gravity of 1.035. Likewise, 0.52 mm, 0.41 mm, 0.36 mm, 0.33 mm, and 0.3 mm should 

be considered as minimum size of granules for specific gravity of 1.01, 1.02, 1.03, 1.04, and 1.05, respectively 

for retention of granules inside the reactor. 

For thick inoculum, suspended solids (SS) > 110 g/l and volatile suspended solids (VSS) to SS ratio < 0.3, 

cationic polymer additives were found to have a negative impact on biomass granulation and COD removal 

efficiency. Based on the statistical models it is concluded that for such thick inoculum, granulation can be 

obtained for treating low strength wastewaters by selecting proper combinations of influent COD and liquid 

upflow velocity, making organic loading rate (OLR) greater than 1.0 kg COD/m 3 .d. 

A Biomass Granulation Index (BGI) has been framed based on mixing in the sludge bed, induced due to 

biogas and superficial upflow velocity of wastewater in UASB reactors. Based upon the above studies, it is 

concluded that good granular sludge (percentage of granules more than 50%, w/w) can be developed in UASB 

reactor if BGI is maintained in the rage of 245 to 480. 

Three kinetic models namely, Monod, Grau second-order, and Haldane model have been studied to predict 

the effluent COD concentration at steady-state. The results indicate that Grau second-order multicomponent 

substrate removal model fits well for estimates of kinetic coefficient in UASB reactors. The investigation of 

nonlinear regression using different error functions as well as prediction analysis reveals association of errors 

with the linearized forms of Monod model used for evaluating the kinetic coefficients. Hence, it is advisable to 

undertake nonlinear regression technique using different error functions in the determination of kinetic 

coefficients for Monod model. 



Microbial community structure in anaerobic degradation of terephthalate and 

phenol 

by CHEN Chia-Lung (ccl9@np.edu.sg) 


Department of Civil Engineering, National University of Singapore 

New PhD Contact: 

Ngee Ann Polytechnic 

Centre of Innovation in Environmental & Water Technology 

Blk 34, #01-01, 535 Clementi Road, Singapore 599489 

+65-64607091 (office) 

+65-96757041 (mobile) 

ccl9@np.edu.sg 

chialung.chen@gmail.com 

Supervisors: 

Assoc. Prof. Wen-Tso LIU 

Department of Civil and Environmental Engineering, University of Illinois at Urbana-Champaign, USA; 

Division of Environmental Science and Engineering, National University of Singapore, Singapore 

Prof. Ong Say Leong 

Division of Environmental Science and Engineering, National University of Singapore, Singapore 

Abstract: 

Microbial community structures in the anaerobic (methanogenic) biological processes treating terephthalate- and 

phenol-containing wastewaters were characterized using 16S rRNA gene-based molecular techniques. The 

overall results showed that the methanogenic microbial communities involved in terephthalate and phenol 

degradation were temperature-dependent, and the main microbial populations were phylogenetically closely 

related. In addition, phenol-degrading enrichment cultures can also degrade terephthalate and benzoate, 

suggesting that these phenol-degrading consortia could be used as the seeding sludge for a bioreactor treating 

terephthalate, or as a co-substrate to enrich terephthalate-degrading microbial consortia, shortening the reactor 

start-up time. Syntrophorhabdaceae (formerly known as Deltaproteobacteria group TA; oval-shaped), 

Pelotomaculum- (fat-rod, some of which may contain spherical spores at the central of cells), and 

Methanosaeta-related cells (filamentous bamboo-shaped or rods with flat-ends) were found as the most 

predominant microbial populations involved in anaerobic terephthalate and phenol degradation. 

List of publications: 

Chia-Lung Chen, Jer-Horng Wu, I-Cheng Tseng, Teh-Ming. Liang and Wen-Tso Liu. Characterization of active 

microbes in a full-scale anaerobic fluidized bed reactor treating phenolic wastewater. Microbes and 

Environments (Accepted, published online 25-Apr-2009). 

Chia-Lung Chen, Jer-Horng Wu and Wen-Tso Liu (2008). Identification of important microbial populations in 

the mesophilic and thermophilic phenol-degrading methanogenic consortia. Water Research 42, 1963-1976. 

Chia-Lung Chen, Hervé Macarie, Ignacio Ramirez, Alejandro Olmos, Say Leong Ong, Oscar Monroy and Wen- 

Tso Liu. (2004). Microbial community structure in a thermophilic anaerobic hybrid reactor degrading 

terephthalate. Microbiology 150, 3429-3440. 



Optimization of fermentation conditions in batch and continuous hydrogen 

production 

by Gustavo Dávila-Vázquez (gdv@ciatej.net.mx) 


Instituto Potosino de Investigación Científica y Tecnológica. División de Ciencias 

Ambientales. Camino a la Presa San José 2055. Lomas 4ª Sección. San Luis Potosí, 

SLP. México. 78216 


Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco. 

Unidad de Tecnología Ambiental. 

Av. Normalistas 800. 

Colinas de la Normal. Guadalajara, Jal. México. 44270 

gdv@ciatej.net.mx 

gdv76@prodigy.net.mx 

Supervisors: 

Dr. Elías Razo-Flores and Dr. Antonio de León-Rodríguez 

Abstract: 

Hydrogen (H 2 ) is seen as a future energy carrier. Because compared to hydrocarbons, it contains 2.75-times 

more energy by weight, besides the fact that its combustion only generates steam as by-product. H 2 can be also 

used for electricity generation with the aid of fuel cells, without the production of greenhouse gases. Around the 

95% of the H 2 used in industry is now produced from hydrocarbons, which makes this process unsustainable. 

On the other hand, biohydrogen (Bio-H 2 ) generated from biomass, is a sustainable energetic alternative which is 

also neutral in greenhouse gases emission. Some microorganisms are capable of produce Bio-H 2 under 

anaerobic conditions, from a wide range of organic substrates. In this thesis, the production of Bio-H 2 was 

studied with glucose, lactose and cheese whey (CW) as substrates, in batch and continuous experiments and 

heat-treated anaerobic granular sludge as inoculum. The effect of initial pH and initial substrate concentration on 

both the hydrogen molar yield (HMY) and volumetric Bio-H 2 production rate (VHPR) was studied for the three 

substrates in batch experiments. Both, higher HMY and VHPR obtained in batch experiments were found at 

higher initial pH than commonly reported as optima in the literature. Furthermore, CW was selected to assess 

Bio-H 2 production with two different media (A and B) due to the high HMY and VHPR obtained, and because 

CW is an industrial by-product. The microbial community analysis showed differences in the microorganisms 

present for the experiments with each medium. Clostridium and Enterobacter species dominated with the use of 

medium B, while experiments with medium A only showed the presence of a proteobacterium. Consequently, 

the use of mineral medium B yielded around twice the VHPR than the obtained with medium A, while HMY 

had a slight increase with the use of medium B. Finally, a continuous stirred tank reactor was operated for 65.6 

days using CW as substrate, supplemented with mineral medium B. Three hydraulic retention times (HRT: 10, 6 

and 4h) were tested attaining the highest VHPR at 6h. Moreover, four organic loading rates (OLR) were 

evaluated at a fixed HRT of 6h. The highest VHPR, 46.61 mmol H 2 /l/h and HMY of 2.8 mol H 2 /mol lactose 

were attained at an OLR of 138.6 g lactose/l/d. The dominant bacterial species at HRT of 10 and 6h belonged to 

the Clostridium genus. The VHPR obtained here in continuous experiments is the highest reported for a stirred 

tank reactor inoculated with anaerobic granular sludge using LS as substrate. The enhancements of both HMY 

and VHPR are critical to assess the full-scale practical application of fermentative processes, which are now 

considered as potential primary generators of sustainable energy in the near future. 


Davila-Vazquez, G., et al. (2008). Fermentative biohydrogen production: Trends and perspectives. Rev Environ 

Sci Biotechnol. 7(1):27-45. http://dx.doi.org/10.1007/s11157-007-9122-7 

Davila-Vazquez, G., et al. (2008). Fermentative hydrogen production in batch experiments using lactose, cheese 

whey and glucose: Influence of initial substrate concentration and pH. Int J Hydrogen Energy. 33(19):4989- 

4997. http://dx.doi.org/10.1016/j.ijhydene.2008.06.065 

Davila-Vazquez, G., et al. (2009). Continuous biohydrogen production using cheese whey: Improving the 

hydrogen production rate. Int J Hydrogen Energy. In press. http://dx.doi.org10.1016/j.ijhydene.2009.02.063 



Operation and modeling of anaerobic sequencing batch reactors (ASBR) in the 

treatment of complex effluents 

by Andrés Donoso-Bravo 


School of Biochemical Engineering, Pontificia Universidad Católica de Valparaíso 


Postdoctoral Researcher 

Department of Chemical Engineering and Environmental Technology 

University of Valladolid. Spain 

Supervisor: 

Dr. Rolando Chamy 

Abstract: 

Anaerobic digestion has become a consolidated and sustainable technology due to its 

economical and environmental benefits. Nowadays, anaerobic digestion is applied in a 

wide variety of organic waste and wastewater and presents comparable advantages over conventional aerobic 

processes. Several anaerobic reactors have been developed, mainly UASB, EGSB and CSTR reactors, however, 

new configurations are being developed and studied to increase the amount of waste treated by anaerobic 

digestion. One of the most important features of the Anaerobic Sequencing Batch Reactors (ASBR) is its 

operational flexibility as a result of its operational variables that may be manipulated such as the reaction time, 

the feeding time or the type of feeding. 

In this work, an Anaerobic Sequencing Batch Reactor (ASBR) has been operated for 8 months to treat low 

strength sewage with high suspended organic matter content simulating domestic wastewater. Three phases of 

operation with increasing Organic Loading Rates (OLR) have been carried out: 0.4 (phase I), 0.8 (phase II) and 

1.2 kg COD/m 3·d (phase III). During the whole experimental period, adequate values of the stability parameters 

(pH, total alkalinity) were obtained. During phases I and II, the removal efficiencies of organic matter 

(expressed as total COD) and total suspended solids ranged between 50 and 60%. However, these values 

decreased to 15-25% in phase III, due to the less HRT the methanogenic biomass was washed out of the reactor 

Afterward, a two-phase anaerobic system using Anaerobic Sequencing Batch Reactors (ASBR) treating the 

same simulated wastewater, to optimize the activity of both populations was implemented. Two reactors in 

series were set, the first one was hydrolytic/acidogenic, while the second one was a methanogenic. Instantaneous 

feeding was used in both reactors. This configuration will promote solid removal reaching higher efficiencies. 

During the experiment, a 69% and 50% of removal efficiencies of total cod removal were obtained for a OLR of 

0.63 and 1.22 kgCOD/m 3·d, respectively. The Solubilized Organic Fraction (SOF) was within a range of 0.3 - 

0.6 gCOD solubilized /gpCOD removed and an average on the Acidified Organic Fraction (AOF) of 0.6 gCOD vfaproduced/gsCOD 

fed , produced in the hydrolytic-acidogenic reactor. These are good results considering the low 

COD influent and the high fraction level of solids content. The methanogenic reactor presented VFA removal 

fraction (VFARF) value between 0.4 – 0.6 gCOD vfa-removed /gCOD vfa-fed for the OLR of 0.63 and 1.22. An overall 

good behaviour of the system was achieved during the whole experimentation, showing that the two-phase 

system was stable, robust and effective for simulated domestic wastewater by anaerobic digestion. 

Then 2 anaerobic sequencing reactors were implemented assessing two operation modalities batch and fedbatch, 

in the phenolic wastewater treatment using an unacclimated biomass was evaluated. It is important to note 

that many industries do not have a continuous discharge of phenol in their effluents, though phenol may be 

released occasionally because of a seasonal change or abnormality in the upstream process. In these cases, it is 

necessary to acquire acclimated biomass or to utilize rapid biomass adaptation in order to treat the effluent. The 

batch reactor showed a poor performance at 5 g/l of cod with a 10, 25 and 40% of the phenol cod, despite a 

readily co-substrate was used in the treatment. A fast adaptation of the biomass was performed in the fed-batch 

reactor using phenol as the only carbon source, reaching a 100% of phenol removal in 10 days. The best results 

were obtained with a 400 mg/l (160 mg/l·d) after 40 days of operation reaching over 95% of phenol removal. 

The phenol concentration impaired the biomass settleability in both reactors, causing an important washout of 

biomass. A fed-batch modalities can be an alternative for the anaerobic treatment of phenolic wastewater. 

Finally, a model with 3 reactions and 2 populations was developed, implemented and validated with the 

experimental data previously obtained. A good correlation between the simulated variables and the experimental 

data was obtained; hence this model might represent a potential tool to control and prediction purposes. 



Study of the effect of process parameters on the thermophilic anaerobic 

digestion of sewage sludge, evaluation of a thermal sludge pre-treatment and 

overall energetic assessment 

by Ivet Ferrer (ivet.ferrer@upc.edu) 

Contact: 

Section of Environmental and Sanitary Engineering 

Department of Hydraulic, Maritime and Environmental Engineering 

Technical University of Catalonia (UPC) 

C/ Jordi Girona 1-3, Builing D1, 08034 Barcelona, Spain 

Tel: +34 93 401 6463 

ivet.ferrer@upc.edu 

Supervisors: 

Xavier Font 

Felícitas Vázquez 

Abstract: 

Energy consumption accounts for some 30 % of the total operating costs of intensive sewage treatment plants 

and, in conventional ones, around 15-20 % of this energy is used in the sludge treatment line. Therefore, 

optimisation of sludge management can contribute in the reduction of wastewater treatment costs. Thermophilic 

anaerobic digestion is more efficient than mesophilic digestion, in terms of biogas production, volatile solids 

(VS) removal and pathogens destruction. The process might be further accelerated by sludge pre-treatment. The 

aim of this PhD Thesis was to study the impact of process parameters on the thermophilic anaerobic digestion of 

sludge, to evaluate the effect of implementing a low temperature pre-treatment step, and to assess alternative 

processes from an energy perspective. 

Two lab-scale reactors were operated for almost two years in order to study the effect of process 

temperature, sludge retention time (SRT), organic loading rate (OLR) and 70 ºC sludge pre-treatment. The 

results were assessed from an energy perspective, by means of theoretical energy balances; and compared to 

those obtained with other experimental data. A first order kinetic model was also used. 

According to the results, methane production rate increased with the OLR (i.e SRT and VS), although 

process unbalance was observed at 6 days SRT (OLR > 5 kg VS m 3 d -1 ). In order to prevent process failure, 

limit values may be: VFA (2.5 g L -1 ), acetate (0.5 g L -1 ), acetate/propionate (0.5), intermediate alkalinity (IA) 

(1.8 g CaCO 3 L -1 ), IA/partial alkalinity (0.9), IA/total alkalinity (0.5), CH 4 (55 %). 

The 70 ºC sludge pre-treatment promoted a rapid sludge solubilization, which was followed by a progressive 

VFA generation after 24 h. Anaerobic digestion of pre-treated sludge (9-48 h) increased biogas production by 

30-40 % working at 55 ºC with a SRT of 10 days. 

Theoretical energy balances suggest that thermophilic sludge digestion would result in net energy production 

provided that digesters with wall insulation and with energy recovery from both the biogas produced and the 

effluent sludge are used. In this case, the energy efficiency would be similar for thermophilic digesters working 

at half the SRT (10-15 days) of mesophilic digesters (20-30 days), meaning that the sludge daily flow rate could 

be increased or the reactor volume reduced, with subsequent savings in sludge treatment costs. Furthermore, 

two-stage systems (70/55 ºC) may result in higher net energy production compared to single-stage systems (55 

ºC) at 10 days SRT. 

Keywords: Anaerobic digestion, energy balance, sludge, thermal pre-treatment, thermophilic. 



Sulfate reducing biological processes in biofilms for metal precipitation 

by Marisol Gallegos García (mgg@ipicyt.edu.mx) 


Instituto Potosino de Investigación Científica y Tecnológica. 

División de Ciencias Ambientales. 

Camino a la Presa San José 2055. Lomas 4ª Sección. 

San Luis Potosí, SLP. México. 78216 

Supervisors: 

Dr. Elías Razo-Flores 

Dra. Mª. Lourdes Berenice Celis-García 

Abstract: 

The environment contamination by heavy metals is an important matter due to its negative impact over the 

public health, the environment and the economy. Sulfate reduction has become and alternative for the treatment 

of wastewaters that contain heavy metals. The general objective of this work was to study the application of 

biological sulfate reduction processes in biofilm reactors for metal precipitation and recovery of the metal 

sulfides in one step. First of all, the competence between methanogens and sulfate reducers was studied in an 

UASB reactor that was operated for over 240 days, for the development of sulfate reducing biomass from a 

methanogenic granular sludge. The chemical oxygen demand (COD) removed by sulfate reduction was 20% and 

the total sulfide concentration reached in the reactor was 310 mg/L. The specific sulfate reducing and 

methanogenic activities of the granular sludge pointed out to the development of sulfate reducing bacteria 

(SRB). The presence of SRB that oxidize incompletely the substrate to acetate, allowed the permanence of 

methanogens that were not limited by substrate, and in consequence were not completely displaced by the SRB 

at the end of reactor operation. Afterwards, a down-flow fluidized bed reactor (DFFB) was proposed for the 

precipitation and recovery of metals. The DFFB reactor was operated continuously during 320 days and 

precipitation efficiencies over 99.0% of Fe, Zn and Cd were obtained. The recovered metal sulfides amounted to 

90% of the theoretical mass expected to be precipitated; pyrite (FeS), sphaleryte (ZnS) and greenockite (CdS) 

predominated in the precipitate. The metal loading rate did not affect the reactor performance even at pH values 

as low as 5.0. The COD removal efficiency was close to 54%, which suggested an incomplete substrate 

oxidation by the immobilized SRB, this was confirmed by the high concentration of acetate in the effluent and 

the absence of alkalinity production. Finally, the development of a sulfate reducing biofilm able to oxidize 

completely the substrate was studied. To achieve this, a selection pressure by substrate was used; three different 

assays were carried out in the DFFB reactor that was fed with a mixture of acetate/lactate with the following 

ratios on a COD basis: 50/50, 80/20 and 90/10 during 21 days. The best biofilm performance in terms of COD 

removal efficiency (71%) and sulfide production rate (2.9 mg H 2 S/L-h) was obtained with the biofilm developed 

in the experiment with the 90/10 acetate/lactate ratio. With this biofilm, the reactor was operated continuously 

110 days. When the pH in the fed was decreased to 4.0, any inhibitory effect was observed over the SRB, as the 

reactor performance showed which achieved COD and sulfate removal efficiencies of 80 and 42%, respectively. 

The alkalinity produced by the complete oxidation of the substrate neutralized the influent pH, and maintained a 

pH of 7.0 within the reactor. This work is the first to report the precipitation and recovery of metals as metal 

sulfides in one step and the development, in a short period of time, of a SRB biofilm able to oxidize completely 

the substrate. 

Link to Thesis: http://www.ipicyt.edu.mx/storage-sipicyt/materialbiblioteca/GallegosGarc%edaMarisol.pdf 


Gallegos-García, M., et al. (2009). Precipitation and recovery of metal sulfides from metal containing acidic 

wastewater in a sulfidogenic down-flow fluidized bed reactor. Biotechnol Bioeng. 102, 91-99. 

doi.wiley.com/10.1002/bit.22049 

Gallegos-García, M., et al. (2008). Competencia por sustrato durante el desarrollo de Biomasa sulfato reductora 

a partir de un lodo metanogénico en un reactor UASB. Sometido, a la Revista Internacional de 

Contaminación Ambiental. 

Gallegos-García, M., et al. (In preparation). Formation of a sulfate-reducing biofilm of complete oxidation of the 

substrate for the treatment of a synthetic effluent with sulfate. 



Characterization and treatment of grey water; options for (re)use 


Sub-department of Environmental Technology 

Wageningen University 

Wageningen, The Netherlands 

Supervisors: 

Jules van Lier 

Grietje Zeeman 

by Lina Abu Ghunmi 

Abstract: 

In addressing the issues of water shortage and appropriate sanitation in Jordan, domestic grey water treatment 

receives growing interest. Grey water comprises the domestic wastewater flows excluding waters associated 

with the toilet. The topics of concern for grey water are its characteristics, treatment options and potentials for 

use after treatment. The overall objective of this thesis is to develop a concept for on-site treating grey water for 

agricultural usage, thus sustaining a recycling process of grey wastewater in arid climate countries, like Jordan. 

A review was made regarding the currently available grey water treatment technologies. In addition, grey water 

was quantitatively and qualitatively characterized, and grey water reuse requirements including treatment, were 

analyzed. Biodegradability and biodegradation rates of the grey water were investigated for selecting appropriate 

design and operation criteria of the treatment technology to be developed. A low-tech semi-technical scale 

treatment system was tested to treat grey water discharges from a dormitory at the Jordan University campus. 

The treatment system was evaluated on obtained removal efficiencies and conformity of the effluent to the 

Jordan guidelines for the use of reclaimed water in irrigation schemes. Finally, the objectives, approaches and 

the results of each chapter are summarized and discussed together with the potentials of applying decentralised 

sanitation and reuse (DeSaR) concepts in Jordan. 

Results show that storage and treatment are prerequisites for any type of grey water use. Grey water is 

aerobically and anaerobically biodegradable but the conversion rates are low. The core of the treatment concept 

consists of an integrated storage and anaerobic treatment unit, fed with a natural influent flow pattern in a downflow 

mode. In the proposed system, up to a one day operational cycle is applied, i.e. a variable HRT 24 hours. 

The second step consists of an aerobic post-treatment, mechanically aerated in a down-flow mode and a one day 

operational cycle, i.e. 24 hours HRT. Both units need insulation in the winter period. The final effluent, stable in 

winter and summer, meets the Jordanian standards for usage in restricted irrigation. The achievable treatment 

efficiency for the CODtot is 44% in the anaerobic unit and 70% in the combined anaerobic-aerobic, unlike the 

high anaerobic and aerobic biodegradability in batch experiments, viz. 70 and 86%, respectively. The highest 

removal efficiency achieved was for the CODss fraction, viz. 71% in the anaerobic and 85% in the combined 

system. Therefore, it is expected that the CODtot removal efficiency of the system can be improved, by 

enhancing the CODcol and CODdis, removal, i.e. applying filtration and/or adding chemicals such as 

adsorbents, coagulants and/or flocculants to the treatment units. 



Combined carbon and nitrogen removal in integrated anaerobic/anoxic sludge 

bed reactors for the treatment of domestic sewage 

by Ghada Nassri Kassab 

Institution: Sub-department of Environmental Technology, Wageningen University, The Netherlands 

Supervisors: Jules van Lier and Bram Klapwijk 

Abstract: The main objective of this research is to assess the applicability and effectiveness of integrating 

anaerobic digestion and denitrification processes in a single sludge system. The integrated concept is of 

particular interest for the treatment of high-strength domestic wastewater and is accomplished by means of 

sequential anaerobic-aerobic systems. The anaerobic pre-treatment can consist of a single anaerobic stage or two 

anaerobic stages, conditioned mainly by the wastewater characteristics, the prevailing ambient temperatures and 

the scale of application. Effluent nitrogen level adjustments using treatment systems consisting of a single 

anaerobic stage followed by an aerobic stage, can be incorporated by circulating part of the nitrified aerobic 

effluent to the influent of the anaerobic stage, integrating denitrification and anaerobic digestion. In the 

sequential treatment system consisting of two anaerobic stages followed by an aerobic stage, nitrogen level 

adjustments can be incorporated by partially circulating the nitrified aerobic effluent to the secondary anaerobic 

stage (methanogenic stage). The first part of this research is focused on studying the potentiality of integrating 

denitrification and methanogenesis processes in single stage UASB reactors operated with flocculent sludge, 

simulating the single anaerobic stage receiving circulated nitrified effluent. Operating a lab-scale UASB reactor 

on synthetic wastewater, simulating high-strength domestic wastewater, under integrated conditions with a 

COD/NO 3 -N ratio of 23 resulted in 92% COD and 97% nitrate removal. Denitrification was the main nitrate 

reduction pathway. Compared to operation under strict anaerobic conditions, methanogenesis reduction upon 

launching the integrated process coincided with reduction of COD available for methane production. Thus, 

indicating that methane production in the UASB reactor was not distressed by the denitrification process 

inhibitory effects. This conclusion is compatible with results obtained from the operation of a semi-technical 

UASB reactor on raw domestic wastewater supplemented with synthetic nitrate. In the latter reactor, integrated 

operation at a COD/NO 3 -N ratio ranging between 20 and 38 didn’t result in significant reduction in observed 

methanogenesis, if compared with operation under strict anaerobic conditions. This preservation of 

methanogenesis can be attributed to two factors. Firstly, under integrated process conditions, an increased 

degree of hydrolysis prevailed that apparently mitigated the reduction in COD available for methane production 

resulting from COD allocation to nitrate reduction processes. Secondly, the sludge specific methanogenic 

activity was preserved under integrated conditions, most likely due to the fact that a substantial fraction of the 

methanogenic biomass was shielded from the oxidized N-compounds in the interiors of the flocculent bio-film. 

Regarding nitrogen removal, nitrate was completely removed. However, it wasn’t completely denitrified, as 33% 

of applied nitrate was reduced via dissimilatory nitrate reduction to ammonium. This obviously decreases the 

efficiency of an integrated system in terms of achieving a high degree of nitrogen removal, at the prevailing 

range of COD/NO 3 -N ratio, i.e. 20-38. Most interestingly, integrated operation didn’t lead to deterioration of the 

flocculent sludge settleability or dewaterability features. The second part of this research is focused on studying 

the integration of methanogenesis and denitrification processes in EGSB reactors for the treatment of pre-settled 

domestic wastewater. The EGSB reactor simulates the secondary anaerobic stage receiving part of the nitrified 

effluent of the succeeding aerobic stage, within a treatment system consisting of two anaerobic stages followed 

by an aerobic stage. Two lab scale EGSB reactors were employed in the study; the first was operated under 

integrated denitrification and methanogenic processes, whereas the second was operated under strict 

methanogenic conditions to set a reference for assessing the impacts of integrated process operation. Moreover, 

the study was carried in two successive periods, in which the applied upflow velocity in the first and second 

periods was 4.5 m.h -1 and 8 m.h -1 , respectively. In the integrated reactor that was operated at a COD/NO 3 -N ratio 

of 20, during the two experimental periods, nitrate was approximately completely removed by means of the 

denitrification process. In the integrated reactor, the COD consumed in methane production was 67% and 69% 

of the COD available for its production during the first and second period, respectively. Such low percentages 

are attributed to “integrated granules” washout of the reactor. This washout is attributed to the growth of 

denitrifiers in form of fluffy biofilm on the surface of granules which in turn deteriorated the granules’ 

settleability. The liquid shear stress induced by the 4.5 m.h -1 and 8 m.h -1 up flow velocities was not enough to 

prevent accumulation of the fluffy biofilm. Of practical interest is the preservation of specific methanogenic 

activity in the sludge grown under integrated conditions. Thus, if the encountered sludge detainment problems 

with integrated granules are resolved, EGSB reactors under integrated conditions can be operated at OLRs 

comparable to those under strict methanogenic conditions. Settleability of integrated granules can be 

substantially improved by periodic application of excessive detachment forces induced either by periodic 

increase in upflow velocity or by periodic circulation of the sludge bed from the bottom to the top of the reactor. 



Research and design of small scale bioreactor 

by Mindaugas Kvasauskas 


Department of Environment Protection, Vilnius Gediminas Technical University, Lithuania 

Supervisor: 

Pranas Baltrnas 

Abstract: 

Due to bad odour, organic waste accumulation facilities should be kept apart from residential areas. A constantly 

increasing amount of organic waste makes it necessary to occupy areas of useful land. A possible technological 

solution to preventing odours and reducing organic waste amounts is using biodigesters for producing gas and 

for further using of the gas to generate energy. Also, anaerobic treatment of organic waste contributes to the 

fulfilment of the Kyoto Protocol requirements for the reduction of greenhouse gas emissions and use of mineral 

fuel resources. 

In order to determine the case when the maximum qualitative and quantitative evolvement of biogas is 

possible under psychrophilic conditions, extintive investigations of evolved biogas were carried out through the 

employment of different biodegradable wastes and their mixtures. 

The psychrophilic temperature (22±2ºC) was maintained in the 6 bioreactors of 220 l in volume. Gas-flow 

and biogas composition (CH 4 , H 2 S, O 2 ) were measured. 

The research was conducted in a number of stages. One-type organic waste was used in the first stage: 

wastewater treatment sludge (WTS), margarine production waste (MPW), fruit and vegetable waste (FV), hen 

manure (HM), pig manure (PM) and grain (G). The second stage employed margarine production waste (MPW) 

mixtures with hen manure (HM) and wastewater treatment sludge (WTS). The third stage used meat waste 

(MW) mixtures with graminaceous waste (GW), wastewater treatment sludge (WTS) and pig manure (PM). The 

fourth stage used mixtures of hen manure (HM) and fruit and vegetable waste (FV), and pig manure (PM) and 

grain (G). The mixtures were made at ratios of 1:3, 1:1 and 3:1 by volume. Prior to the experiment, a substrate 

containing 10% of dry substance of its total mass was prepared. 

It was determined experimentally that the highest biogas production rate under psychrophilic conditions 

when treating biodegradable waste of one type can be obtained using pig manure, 33 l/m 3 d, and the smallest – 

treating margarine production waste, 14 l/m 3 d. The highest concentration of methane was recorded when using 

pig manure (87.8%) and wastewater treatment sludge (86.4%), whereas the lowest – when using margarine 

production waste (4.8%). The highest biogas production rate under psychrophilic conditions when treating 

biodegradable waste of two types can be obtained using a mixture of meat waste and pig manure 1:3, 34 l/m 3 d, 

and the smallest – treating a mixture of margarine production waste and wastewater treatment sludge 3:1, 8 

l/m 3 d. The highest concentration of methane was recorded when using a mixture of pig manure and grain 3:1, i. 

e. 87.8%, whereas the lowest – when using a mixture of margarine production waste and poultry manure 3:1, i. 

e. 4.8%. Also it was established experimentally that the optimum period of keeping biomass in the bioreactor 

under psychrophilic conditions is from 2 to 3 weeks. 

Only the data when CH4 concentration in biogas reached 65% are given. The total energy value in kJ, recorded 

during the days of the experiment, when biogas was suitable for burning, as well as the medium energy value. 

The greatest amount of energy can be produced during anaerobic treatment of organic waste under 

psychrophilic conditions from PM. The total energy value reaches 4696 kJ from 1 m 3 biomass per experiment 

time when CH 4 concentration reached 65%, and the medium value – 294 kJ/m 3 . Good results were also achieved 

when using a PM and GW 3:1 mixture. The total energy value reaches 3278 kJ from 1 m 3 biomass per 

experiment time when CH 4 concentration reached 65%, and the medium value – 234 kJ/m 3 . 

The lowest energy value under psychrophilic conditions was obtained when using MPW and WTS 1:3. The 

total energy value reaches 1376 kJ (58% less than in the case of PM) and the medium value – 153 kJ/m 3 (35% 

less than in the case of PM). 

To intensify biogas production process, mezophylic or thermophilic temperature should be held in 

bioreactors. The improved bioreactors are suitable for efficient processing of different solid and liquid 

biodegradable wastes generating in agriculture, food industry and wastewater treatment plants. 



Improving stability of anaerobic digesters for animal waste treatment 

by Marcelo Loureiro Garcia (mlgarcia@rc.unesp.br) 


Assistant Professor 

Universidade Estadual Paulista (UNESP) 

Instituto de Geociências e Ciências Exatas de Rio Claro 

Departamento de Petrologia e Metalogenia 

Av. 24/A, 1515. Bela Vista 

Rio Claro, SP 13506-900, Brasil 

Tel: +55 (19) 3526 2833 

mlgarcia@rc.unesp.br 

Supervisor: 

Largus (Lars) Angenent, PhD 

Associate Professor 

Biological and Environmental Engineering 

Cornell University 

214 Riley-Robb Hall 

Ithaca, NY 14853, USA 

Tel: +1-607-255-2480 Fax: +1-607-255-4080 

la249@cornell.edu 

Abstract: 

In countries where concentrated animal feeding operations (CAFOs) are used, implementation of animal waste 

treatment is desired for both environmental protection and energy recovery. The capability of anaerobic systems 

to produce large quantities of biogas from animal wastes has been demonstrated, however, a lack of knowledge 

on the microbial community structure exits. Four 5-liter anaerobic sequencing batch reactors (ASBRs) were 

operated over a period of 988 days to treat swine waste. Over the course of three distinct operating periods, the 

performance of the bioreactors was assessed with conventional environmental engineering techniques, while 

archaeal and bacterial microbial communities were unraveled with molecular biology techniques. The bacterial 

community analyses did not show a significant difference between low- and high-ammonia reactors for different 

temperatures at the phyla level, with Firmicutes and Bacteroidetes as the predominant bacterial phyla. Members 

of the family Methanosarcinaceae and of the order Methanomicrobiales predominated as acetoclastic and 

hydrogenotrophic methanogens, respectively. The performance of the anaerobic digesters was clearly affected 

by varying total ammonium concentrations and mesophilic temperatures, however, this did not result in a 

considerable microbial community shift. Carbon and energy sources outplayed toxicity effects of ammonia and 

temperature constraints of kinetics and thermodynamics in determining the microbial bioreactor communities. 

However, some changes in the community were found. For instance, a gene sequencing survey from a sample 

enriched with 13 C-acetate indicated an enhancement of sequences from the genus Pseudomonas (among other 

uncultured phyla), explaining the 25% methane that was generated via an alternative pathway: syntrophic 

acetate oxidation (with GC-MS). This pathway only occurred at very high ammonia concentrations (5,200 total 

ammonium-N (mg/L)), which prevented unstable conditions in the bioreactors with high ammonia levels. We 

showed that powerful molecular biology techniques are needed to monitor incremental changes in the 

community structure that can have large effects on reactor stability. 



Mesophilic Fermentative Hydrogen Production from Sewage Biosolids 

by Jaime Massanet-Nicolau (jmassane@glam.ac.uk) 


Sustainable Environment Research Centre (SERC). 

Faculty of Health, Sport and Science 

University of Glamorgan 

Pontypridd, CF37 1DL, UK 

Supervisor: 

Prof Richard Dinsdale 

Prof Alan Guwy 

Prof Andrew Wheatley 

Abstract: 

The increasing cost of fossil fuels, combined with concerns about their impact on our environment has led to a 

renewed interest in hydrogen as a clean, sustainable, alternative energy vector. Using sewage biosolids as the 

substrate for fermentative hydrogen production offers several advantages over the use of other biomass sources. 

It is available at little or no cost and is abundant, being produced wherever there are human settlements, with 1.3 

million tonnes (dry solids) per year currently being produced in the U.K alone. 

This research demonstrated the feasibility of hydrogen production from sewage biosolids via anaerobic 

fermentation. To do this a number of issues specifically relating to the nature of sewage biosolids had to be 

addressed. Firstly, the solids content and rheology made automatic feeding difficult. The feedstock also 

contained high levels of indigenous microorganisms and a high ratio of insoluble to soluble carbohydrate. 

To address these challenges, a novel reactor design using wide bore tubing and computer controlled pumping 

equipment was successfully used to construct a working continuously fed bio-reactor. A combination of heat 

treatment at 70 o C for one hour and pre-treatment with a commercially available food processing enzyme mixture 

was found to be the most efficient method of inactivating competing microorganisms and improving substrate 

quality. 

Hydrogen was successfully produced via batch fermentation of primary sewage biosolids which had 

undergone heat treatment and enzymatic digestion. When fermentation took place at pH 5.5 a peak hydrogen 

production rate of 3.75 cm 3 min -1 was observed. At this pH the hydrogen yield was 0.37 mol H 2 mol -1 

carbohydrate, equivalent to 18.14 L H 2 kg -1 dry solids. Fermentative hydrogen production from sewage biosolids 

was also demonstrated in a five litre, continuously fed bio-reactor for the first time. A comparison of different 

hydraulic retention times showed that hydrogen production was most stable at a HRT of 24 hours. A hydrogen 

producing fermenter was successfully linked to a methanogenic bio-reactor in a two stage digestion process. 



Anaerobic digestion in sustainable biomass chains 


Sub-department of Environmental Technology 

Wageningen University 

Wageningen, The Netherlands 

Supervisors: 

Jules van Lier 

Rudy Rabbinge 

by Claudia P. Pabon Pereira 

Abstract: 

This thesis evaluates the potential contribution of anaerobic digestion (AD) to the sustainability of biomass 

chains. Results provide insights in the technological potential to recover energy and valuable by-products from 

energy crops and residues, and evaluate biomass cascades involving AD technology for their feasibility and 

desirability. Embedding AD in biomass chains addresses current constraints towards increased use of biomass 

for energy production considering land competition and environmental pollution. Within this panorama so far 

major advantages of AD to improve energy efficiency and closing material cycles have received a limited 

attention. As part of the experimental research an Oxitop® protocol was refined for screening plant material 

suitable for anaerobic digestion based on their energy content. Environmental factors influencing the test 

outcome are the use of NaOH pellets for CO 2 scavenging, substrate pretreatment, microbial culture, and type of 

buffer. The use of NaOH pellets and substarte pretreatments were most influential on the results. By means of 

the developed Oxitop® protocol the relationship between plant ligno-cellulosic composition and the biomethane 

potential (BMP) and hydrolysis rates was researched. The Acid Detergent Fibre (ADF) and the Neutral 

Detergent Fibre (NDF) as analyzed by the van Soest method were proposed as suitable plant characterization 

techniques for predicting the biodegradability and hydrolysis constants. The biodegradability model developed 

was further used to predict the biodegradability of 114 European plant samples identifying interesting crops and 

crop residues suitable for anaerobic digestion. Batch experiments on digestate quality showed an increase of 20- 

26% and 0-36% in solublised NH 4 + and PO 4 3- , respectively, after 2 months of digestion. The largest fractions of 

the inorganic nutrients were found in the liquid fraction of the digestate, i.e. 80-92% NH 4 + and 65-74% PO 4 3- . 

Increase in manure content in the mixture showed a positive effect in the methane production rate and the total 

amount of nutrients in the digestate. 

The added value of AD within different biomass cascades was evaluated using the sustainable development 

concept in a specified system boundary. The sensitivity analysis of the energy balance of an AD facility shows 

that heat losses become the most important energy loss when high value substrate such as energy maize are 

employed. In contrast, when a low energy substrate such as manure is used, the indirect energy of the 

infrastructure facilities, become the most important energy input. A sustainability framework was proposed for 

evaluating the potential role of AD in biomass cascades applied for the Colombian situation. Results show that 

production of bio-ethanol from cassava is only sustainable from an energy and greenhouse gas (GHG) 

perspective when energy recovery from the process residues, using AD, is part of the process. The exact 

outcome of the evaluation largely depends on variables like substrate drying, type of fuel used and type of 

applied AD system. During the study of other biomass cascades the contribution of by-products was shown to be 

crucial, constituting 41-68% of the sum of all energy flows. For oil palm, sugarcane, panelacane and cassava, 

the energy contribution of the byproducts to the different biofuel systems fluctuates between 51-158, 122-290, 

71-170, and 36-71 GJ.ha -1 yr -1 , respectively. AD had also a positive impact on nutrient recovery and water 

savings in the studied chains. The energy, nutrient and water benefits were set in perspective by giving an 

indication on the economic benefits and land savings potentially attainable under Colombian conditions. 



Modeling Microbial Diversity in Anaerobic Digestion 

by Ivan Ramírez (ramirezy@supagro.inra.fr) 


Laboratoire de Biotechnologie de l'Environnement 

LBE-INRA Avenue des étangs - 11100 Narbonne, France 

Tel: +33 468.425.151 - Fax: +33 468.425.160 

ramirezy@supagro.inra.fr 

Supervisor: 

Dr. Jean-Philippe Steyer 

Laboratoire de Biotechnologie de l'Environnement 

LBE-INRA Avenue des étangs - 11100 Narbonne, France 

tel: +33 468.425.151 - Fax: +33 468.425.160 

steyer@supagro.inra.fr 

web: http://www.montpellier.inra.fr/narbonne 

Abstract: 

This new century addresses several environmental challenges among which distribution of drinking water, 

global warming and availability of new in substitution of fossil fuels are of the most importance. Among other 

renewable sources, biogas production from wastes is particularly interesting. Moreover, a techno-economical 

comparison demonstrated the benefits of a two-step process (H 2 +CH 4 ) compared to the classical one-step 

methane production. Economic evaluation of biogas plants has revealed that many plants can only survive 

economically if special incentives are applied. Nowadays, it is thus necessary to find ways to optimize the 

biogas production in order to make biogas plants economically viable with decreased or no subsidies. 

Optimization of the biogas process by advanced monitoring and control can undoubtedly lead to better economy. 

Such strategies require, in general, the development of appropriate mathematical models, which adequately 

represent the main biological processes that take place. Moreover, experimental evidence is available suggesting 

that the structure and properties of a microbial community may be influenced by process operation and on their 

turn also determine the reactor functioning. In order to adequately describe these phenomena, more detailed 

microbial diversity should be taken up in mathematical models. This was demonstrated in this work by 

extending the ADM1 model to describe microbial diversity between organisms of the same functional group. 

The developed model was further shown useful in assessing the relationship between reactor performance and 

microbial community structure in different conditions: difference in reactor configurations, influents and load 

regimes, both in normal and abnormal situations. As engineered systems are often more manageable than largescale 

ecosystems, and because parallels between engineered environments and other ecosystems exist, we 

showed in this work that the former was used to elucidate some unresolved microbial ecological issues. 



Bioaugmentation for recovery of anaerobic digesters subjected to a toxicant 

by Anne E. Schauer-Giménez 


Marquette University, December 2008 


Post-doctoral Fellow 

Marquette University 

Water Quality Center 

Milwaukee, WI (USA) 

Supervisor: 

Dr. Daniel H. Zitome 

Abstract: 

Traditionally, practitioners have considered anaerobic methanogenic cultures as “black boxes” and have not paid 

considerable attention to the microorganisms. However, the methanogenic community is important because 

different populations may behave differently in terms of methane production rate or stability during process 

upset. The research presented in this dissertation explored the diversity of Bacteria and Archaea communities in 

various methanogenic enrichment cultures. 

With the help of molecular techniques (PCR, cloning, restriction digests, and sequencing), Bacteria and 

Archaea clone libraries were constructed for four enrichment cultures herein referred to as C1, C2, C3, C4. The 

enrichment conditions differed based upon substrates and oxygen fed, and were as follows: C1 – H 2 :CO 2 , C2 – 

H 2 :CO 2 + glucose, C3 – H 2 :CO 2 + air, and C4 – H 2 :CO 2 + glucose and air. It was hypothesized that the varied 

conditions would have an effect on microbial community diversity. 

When comparing the enrichment cultures that received air versus those that did not, a shift in methanogen 

community occurred. In C1 and C2, Methanosaeta was the most dominant genus. On the other hand, 

Methanosaeta was not dominant in C4 and not found in any sequences in C3. Methanospirillum was the most 

common methanogen genus in C3. These results indicate that addition of air had an effect on the methanogen 

community. 

Bioaugmentation, the addition of a specific microorganism or mixed culture to enhance a desired activity 

(e.g. methane production or COD reduction), can potentially be used to increase the recovery rate of anaerobic 

wastewater treatment facilities after exposure to toxicants. In the work describe herein, cultures C2 and C4 were 

used to bioaugment upset anaerobic digesters. The bioaugmentation investigations successfully demonstrated 

the ability of the bioaugmentation culture to increase methane production and recovery rate of a stressed digester 

exposed to the toxicant, oxygen. Methane production increased an average of 47% in the bioaugmentation 

digesters as compared to the controls, which were not bioaugmented. Also, SCOD concentrations in the 

bioaugmented digesters decreased below 2,000 mg/L 83 days sooner than the controls. Bioaugmentation is a 

promising approach to help decrease the recovery time of anaerobic digesters exposed to a toxicant. 



Effects of Macrolide Antimicrobials on Anaerobic Treatment Systems 


University of Illinois at Urbana-Champaign (USA) 

Supervisors: 

Lutgarde Raskin (advisor) 

Julie Zilles (co-advisor) 

Eberhard Morgenroth (co-advisor) 

by Toshio Shimada (TShimada@carollo.com) 

Abstract: 

Antimicrobials are used commonly in animal production facilities for treatment and prevention of disease and as 

growth enhancers. An anaerobic sequencing batch reactor (ASBR) was operated with a synthetic, carbohydratebased 

wastewater to study the effects of tylosin, a macrolide antimicrobial commonly used in swine production, 

on treatment performance, microbial community structure, and development of antimicrobial resistance. The 

experimental period was divided into three consecutive phases with different influent tylosin concentrations (0, 

1.67, and 167 mg/L). The addition of 1.67 mg/L tylosin to the reactor had negligible effects on total methane 

production and effluent chemical oxygen demand (COD), yet analyses of individual ASBR cycles revealed a 

decrease in the rates of methane production and propionate uptake. The addition of 167 mg/L tylosin to the 

reactor resulted in a gradual decrease in methane production and the accumulation of propionate and acetate. 

Batch tests indicated that the specific biogas production (SBP) with the substrate butyrate was completely 

inhibited in the presence of tylosin. Measurements of treatment performance together with phylogenetic and 

quantitative fluorescence in situ hybridization (FISH) analyses suggested that the addition of 1.67 mg/L and 167 

mg/l of tylosin inhibited homoacetogenic bacteria, propionate oxidizing syntrophic bacteria, and aceticlastic 

methanogens. Tylosin directly inhibited these two bacterial populations, while inhibition of the methanogenic 

population was presumed to be indirect and was attributed to a decrease in the pH of the reactor. 

In addition to evaluating the impact of tylosin on anaerobic treatment, the accumulation and degradation of 

storage compounds was studied in the glucose-fed ASBR. Total carbohydrate analysis showed that 26% of the 

glucose added to the reactor transiently accumulated within the biomass. 13C nuclear magnetic resonance 

(NMR) analysis identified the main carbohydrate produced as trehalose (alpha-D-glucopyranosyl-(-Dglucopyranoside)). 

The International Water Association Anaerobic Digestion Model No. 1 (ADM1) was 

modified to include microbial storage and hydrolysis of reserve carbohydrates, and tylosin liquid-solid mass 

transfer and inhibition. This modified model adequately described accumulation and degradation of storage 

compounds as well as the microbial population dynamic profiles observed in the ASBR. The modeling results 

confirmed that tylosin inhibited saturated fatty acid beta-oxidizing syntrophic bacteria and suggested inhibition 

of fermentative bacteria resulting in unfavorable effects on methanogenesis. 

Keywords:: ADM1, anaerobic, macrolide, methanogenesis, storage, SBR, tylosin 



Development of High-Rate Anaerobic Granular Sludge Bed Process for Low 

Strength Wastewater Treatment at Low Temperature 

by Wilasinee Yoochatchaval (wilasinee.y@nies.go.jp) 


Post doctoral researcher, Water and Soil Environment division, 

National Institute for Environmental Studies, 16-2 Onogawa, 

Tsukuba, Ibaraki, Japan 305-8506 

Email: wilasinee.y@nies.go.jp, wilasinee_y@hotmail.com 

Supervisors: 

Dr. Kazuaki Syutsubo, Senior Researcher of National Institute for Environmental 

Studies, Japan 

Dr. Takashi Yamaguchi, Associate Professor of Nagaoka University of Technology, 

Japan 

Dr. Hideki Harada, Professor of Tohoku University, Japan 

Abstract: 

In this study, the anaerobic granular sludge process and its operation mode were established for efficienttreatment 

of the low strength wastewater at low temperature. First of all, the feasibility of an expanded granular 

sludge bed (EGSB) reactor for low strength wastewater with 0.6-0.8 g COD/L was tested at 20°C. In this 

process, methanogenic granular sludge was used as seed and recirculation of effluent was made to fluidize the 

sludge bed. As a result, sludge retention in this process was very well (over 40 days). Consequently, this process 

was possible to achieve high rate treatment of low strength wastewater at hydraulic retention time of 1.5 h with 

COD loading rate of 12.8 kg COD/m 3 /day. 

The influence of decreasing in the temperature and the organic concentration of the wastewater was 

evaluated in lab-scale EGSB reactor. The reactor exhibited sufficient COD removal efficiency until 10°C 

operation due to the good-microbial adaptation of granular sludge at low temperature. The significant increase 

of hydrogen-fed methanogenic activity between test temperatures of 15°C and 20°C was observed. At this time, 

proliferation of psychrotolerant methanogen (hydrogen utilizing Methanospirillum spp.) in the retained sludge 

was confirmed. On the other hand, decrease of organic concentration of wastewater (from 0.8 to 0.4 gCOD/L) 

has caused the deterioration of organic removal efficiency and physical property of retained sludge due to the 

inactivation of anaerobic bacteria. To solve this problem, new operational technology was proposed. Control of 

ORP of influent and intermittent the effluent recirculation are possible to active the microbial activity of 

methanogens in the retained sludge. 

To clarify the effect of sucrose content of fed-wastewater on the physico-microbial characteristics of 

granular sludge, we operated two reactors at 20°C by sucrose based wastewater (sucrose, 90% of COD) and 

volatile fatty acid based wastewater (sucrose, 0% of COD). Sucrose founded much negatives effects on the 

granular sludge physical properties and microbial properties (bulking and wash out of retained sludge due to the 

overgrowth of acidogens) at low temperature condition. 



ONGOING ANAEROBIC DIGESTION PROJECTS 

The Wales Centre of Excellence for Anaerobic Digestion was established in 2008 at the University of 

Glamorgan, Wales, UK. The Centre is Directed by Dr Sandra Esteves and was created to assist with the creation 

of an infrastructure of AD plants in Wales to treat municipal food wastes and other biodegradable commercial 

and industrial wastes. 

Building on the considerable research experience developed over the past 40 years in the Sustainable 

Environment Research Centre at the University of Glamorgan, the Centre of Excellence represents an effective 

means of using this expertise to support the growth of an AD industry in Wales and the UK. 

The Centre of Excellence has a mandate to provide two key roles in Wales. Firstly the Centre is working 

towards removing barriers to implementing AD caused by a lack of understanding of the technology. This will 

be achieved through a programme of awareness raising and provision of impartial information and advice. The 

Centre has already engaged with key policy and regulatory stakeholders including local authority planning 

authorities, the Environment Agency and government procurement teams. 

Secondly, the Centre of Excellence is providing technical assistance and guidance to those directly involved 

in establishing AD plants within Wales. This ranges from pre-feasibility discussions, delivery of feasibility 

studies through to substrate and digestate characterisation, biogas potentials and longer term pilot scale studies 

undertaken at the Centre’s laboratory facility. 

Centre Director Dr Sandra Esteves says “Anaerobic digestion will, over a 

relatively short timescale, become a key waste management and bio-energy 

generation technology in Wales and the UK. The Wales Centre of Excellence for 

Anaerobic Digestion has been created to help those implementing, regulating 

and utilising the technology to maximise the environmental and economic 

benefits to Wales. The Centre should ensure that this is achieved through the 

development of an integrated and technically robust infrastructure and a long 

term plan to support the management of these facilities once in place.” 

Funding to establish the Centre has been provided by the Welsh Assembly 

Government. 

For further information please contact: 

Dr Sandra Esteves (sesteves@glam.ac.uk) Tel. +44(0)1443 624130. 

or visit: www.walesadcentre.org.uk 



The PROBIOGAS Project 

The objective of the PROBIOGAS project is to develop sustainable systems of production and use of biogas in 

agro-industrial areas, the demonstration of its viability and its promotion in Spain. The project has been 

considered by the Ministry of Science and Innovation both singular and strategic, having received funds from 

the Ministry itself and from the European Regional Development Fund (ERDF). The project began at the end of 

2007 and will finish in 2010. The project has a total eligible budget of 6.7M€ and has received economical 

support like grants and refund anticipations loans. 

The activities of this project are centered in biogas obtained from wastes of agro-industrial origin (animal 

farms, agriculture, agro-industry, food industries, bioenergy industries, etc.); landfill gas and biogas originating 

from urban wastewater purification stations are not considered. 

In addition, all the studied techniques of anaerobic digestion in the project always apply the concept of "codigestion" 

(joint treatment of organic wastes of different origin and composition). In PROBIOGÁS the codigestion 

of combinations of the most important agro-industrial wastes in Spain will be studied. 

The project consists of twelve inter-related subprojects, which 

have their own specific objectives, although they share the common 

objective of PROBIOGAS. Thus, part of these subprojects will 

involve an investigation into the improvement of the production 

systems of biogas based on the anaerobic co-digestion of the more 

frequent agro-industrial wastes in Spain. Others will investigate ways 

of improving the agronomic use of the digestate in traditional and 

power cultures. Also, a study of the viability of the possible uses of 

biogas in vehicles, facilities of co-generation, microturbines and fuel 

cells, as well as their introduction in natural gas networks will be carried out. In addition, seven actions of 

technological demonstration including innovating actions will be made as much in biogas production techniques 

as in the use of the digestate and biogas. 

In this project there is ample participation (28 partners), divided in companies (15) and research centers (13) 

and which, although they develop their activity in different sectors, share their interest in the advantage of 

production and use systems of biogas. 

The relation of partners is as follows: ABANTIA, Energía y Medio Ambiente (AEMA), AINIA Centro 

Tecnológico, Purines Almazán S.L., Biogas Fuel Cell (BFC), CSIC-CEBAS, CESPA, Fundación CIDAUT, 

Centro de Investigaciones Energéticas Medioambientales y Tecnológicas (CIEMAT), SAC Ganadera del Valle 

de los Pedroches (COVAP), Fundación Asturiana de la Energía (FAEN), Fundación Patrimonio Natural de 

Castilla y León (FNAT), Centro Tecnológico de Gestión Integral de Residuos Orgánicos (GIRO), S.A.T. San 

Ramón (GSR), GUASCOR, Instituto para la Diversificación y Ahorro de la Energía (IDAE), Universidad de 

León (IRENA UL), Instituto Valenciano de Investigaciones Agrarias (IVIA), NATURGAS ENERGÍA S.A., 

PROTECMA, FUNDACIÓN RURAL CAJA, Sociedade Galega do Medio Ambiente (SOGAMA), Unión 

Temporal de Empresas TETMA S.A. -URBASER S.A., Universidad de Barcelona (UB), Universidad de Cádiz 

(UCA), Universidad Miguel Hernández (UMH), Universidad de Oviedo (UNIOVI), Universidad Politécnica de 

Valencia (UPV) and Universidade de Santiago de Compostela (USC). 

The coordinator of the project is AINIA, in the person of Mr. Andrés Pascual. For more information, see Web 

page: www.probiogas.es 



ANAMIX – Assessment of rheological behavior and mass transfer properties for the 

improvement of dry digestion processes 

Methane fermentation is a biological process converting organic matter into a methane-rich gas (biogas). It is 

mostly used for municipal and agricultural organic wastes, which constitute a valuable resource: the European 

production of biogas was equivalent to 5 million tons of oil in 2006. Most systems used for municipal wastes 

today are called dry digestion processes: they are more efficient and less water-consuming than classical 

systems, but the mechanical structure of the digestion medium is still ill-defined and difficult to control at the 

industrial scale. The essence of the ANAMIX project is to answer to specific questions associated to dry 

digestion, using new techniques and new approaches for the description of dense and pasty media. It combines 

Process Engineering (rheology, mass transfer, mixing) and Life Science (microbial kinetics and ecosystem 

ecology) together to study the complex interactions occurring in dry digestion, and to propose a model enabling 

the simulation of the mechanisms. Our project is guided by a double expectation: first, to improve our 

knowledge of basic mechanisms that take place in dry digestion systems; Second, to provide the tools for 

making the future industrial systems easier to design and to operate. 

Partnership: 

INSA Lyon, Laboratory for Civil and Environmental Engineering (http://lgcie.insa-lyon.fr/index.phpRub=1) 

INRA Narbonne, Laboratoire de Biotechnologie de l’Environnement 

VALORGA INTERNATIONAL 

Coordinator: Pierre BUFFIERE – INSA de Lyon (pierre.buffiere@insa-lyon.fr) 

Financial support: 528,841 €, funded by the french National Research Agency (ANR), Bioenergy Call 2008. 

Dates: From December 2008, 36 months 

Metal speciation in anaerobic sludge digesters supplemented with trace nutrients to 

enhance biogas production 

School of Civil Engineering and School of Geography, Earth and Environmental Sciences, 

University of Birmingham, UK 

Supervisors: Dr CM Carliell-Marquet; Dr John Bridgeman; Dr Joanna Renshaw 

PhD students: Farryad Ishaq; Jimmy Roussel 

Project sponsors: Engineering and Physical Sciences Research Council (UK) and Severn Trent Water Ltd. 

Anaerobic sludge digesters are metal-rich environments. Metals coming into the digesters are rapidly 

precipitated as carbonates, sulphides and, to a lesser extent, phosphates. Methanogenic bacteria have specific 

requirements for metals such as nickel, cobalt and iron. Research has shown that supplementing anaerobic 

digesters with trace amounts of these required metals can increase the methane yield. Other research has 

highlighted the fragile balance that exists in these digesters between beneficial cations and anions (e.g. iron and 

phosphate). For example, iron supplementation can increase methane production but excess iron can cause 

phosphate to be precipitated, rendering it biologically unavailable, which could in turn decrease methane 

production. 

This introduces two key themes about which little are understood. Firstly, the issue of bioavailability: In what 

form is a metal bioavailable in an anaerobic digester In what form is a metal biologically unavailable 

Secondly, the issue of chemical equilibrium, i.e. that every change made to the inorganic balance of an anaerobic 

digester has multiple knock-on effects to cations and anions within the digester. Hence, the true effect of adding 

a metal supplement to increase methane yield is difficult to predict. 

This research investigates the effect of trace metal supplementation on biochemical reaction rates and metal 

speciation in anaerobic sludge digesters. Controlled laboratory digestion experiments, e.g. acetate utilisation rate 

or hydrolysis rate tests, are being used to measure the effect of trace nutrient supplements on different stages in 

the digestion process. Metal fractionation experiments on digesters and digested sludge, combined with 

equilibrium speciation models, are being used to improve our understanding of metal speciation and 

bioavailability in laboratory digesters supplemented with trace nutrients. An important outcome of this research 

is determining solubility products for compounds in a digested sludge matrix, as opposed to pure water, so that 

existing equilibrium speciation models (eg. MINTEQA2) can be used to reliably predict speciation scenarios in 

anaerobic sludge digesters. 



200 WTPD Thermal Dryer Using Landfill Gas as the Primary Fuel Goes Online In 

Manatee County Florida 

By: Alex L. Monroe, PE, Sr. Engineer/Scientist McKim & Creed, PA, 104 Gillespie Street, Fayetteville, NC 

28301 USA, (E-mail: amonroe@mckimcreed.com) 

INTRODUCTION 

Manatee County is located on the west coast of Florida and, like so many other areas in Florida, is experiencing 

rapid growth, which at times taxes the capacity of its wastewater utility infrastructure. The county owns and 

operates three water reclamation facilities: the North, the Southeast and the Southwest. All have non-discharge 

permits (total current capacity is 40.5 MGD). All the wastewater treated by the county’s WRFs is land-applied 

on residential sites, farmlands or golf courses. The following lists the dryer’s fuel requirements for the dryer 

purchased by the county. 

Table 1 Biosolids Fuel Requirements 

Dryer Fuel Natural Gas Landfill 

Gas 

High Heating Value, BTU/SCF 1,038 410 

Fuel Volume, CFH 19,269 48,700 

Dryer, Therms/hr 200 202 

Total Therms/hr (with 10% Contingency) 220 222 

Since the landfill has an abundance of methane gas, the 

landfill gas was selected as the primary fuel while natural gas 

was selected as the backup fuel for the dryer. The county 

anticipates that it will save up to $1.7 million per year by using 

landfill gas as the primary fuel for the dryer. 

CONCLUSIONS AND LESSONS LEARNED 

The project has been online since August of 2008 and is producing Class AA pellets of exceptional quality. 

The main problem concerning the landfill gas was high carbon dioxide and nitrogen emissions from the 

dryer that were in violation of the Clean Air Act. To overcome this problem, the county elected to install a 

thermal oxidizer to treat the emissions from the dryer. Natural gas had to be used to power the thermal oxidizer 

due to the variability in the quality of the landfill gas since the landfill gas was not scrubbed to remove 

contaminants. Another problem that had to be overcome due to the varying quality of the landfill gas resulted in 

natural gas having to be used with the landfill gas to provide optimum drying capacity. The ratio of landfill gas 

to natural gas turned out to be 70 percent landfill gas and 30 percent natural gas. This reduced the anticipated 

savings in the cost for natural gas. 

The biosolids generated by the county is meeting the State of Florida’s Class AA biosolids criteria following 

the drying process. The county intends to use the pellets from the dryer as a soil amendment for the daily cover 

at the landfill, and plans to sell the pellets to local farms and residents as a fertilizer. The savings over natural 

gas will also provide the county a payback for the capital cost of the dryer over an eight- to ten-year period. 



ANAEROBIC URL SITES 

Teaching, awareness and training sites 

www.uasb.org 

Anaerobic knowledge centres: 

www.ftns.wau.nl/lettinga-associates/ 

www.walesadcentre.org.uk 

www. orstom.fr/loma 

www.enzyme.chem.msu.ru/ekbio/eng/index.html 

Anaerobic digestion companies 

3R Ingeniería Ambiental SL 

ADI 

Aquatec Maxcon 

Biotec 

Biotecs 

Biothane Systems International 

Biotim 

Ecomembrane 

Ecovation 

Entec Biogas GmbH 

Global Water Engineering 

Grontmij 

IBTech 

NewBio, Inc 

Ondeo Degrémont 

Paques 

Proserpol 

Purac 

Rains 

Reva Enviro 

http://www.3r-ingenieria.es 

http://www.adi.ca 

http://www.aquatecmaxcon.com.au 

http://www.bio-tec.net 

http://www.biotecs.com.br 

http://www.biothane.com 

http://www.biotim.be 

http://www.ecomembrane.com 

http://www.ecovation.com 

http://www.entec-biogas.at 

http://www.globalwaterengineering.com 

http://www.grontmij.nl 

http://www.ibtech.com.mx 

http://www.newbio.com 

http://www.degremont.fr 

http://www.paques.nl 

http://www.proserpol.com 

http://www.lackebywater.se/en 

http://www.rainsgroup.com 

http://www.revaenviro.com/index.htm 

Other sites with AD links: 

http://www.ad-nett.org/

Specialist Group on ANAEROBIC DIGESTION Newsletter - IWA

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