1. Use the left mouse button to move forward through the show Use the right mouse button to view the slides in normal view, edit or print the slides The following slides are provided by Dr. Vincent O’Flaherty.

2. High-rate reactor designs Anaerobic digester designs based on biomass retention: (a) anaerobic filter/fixed bed reactor; (b) downflow stationary fixed-film reactor; (c) expanded bed/fluidised bed reactor; (d) upflow anaerobic sludge blanket reactor; Expanded granular Sludge Bed (e) hybrid sludge bed/fixed bed reactor

3. USE OF ANAEROBIC DIGESTION FOR INDUSTRIAL WASTEWATER TREATMENT Installation of anaerobic digesters for industrial wastewaters has grown very rapidly over the past 15-20 years. UASB design is the most widely used, EGSB becoming more common. Very high loading rates and biogas productivity; HRT typically 1 day or less.

4. Up to 30 kg COD/m 3 /d - UASB; 100 kg COD/m 3 /d - EGSB Up to 20 m 3 biogas/m 3 /d Typically achieve 80-99% COD removal. A.D. treated wastewater is either discharged to the municipal sewer for final treatment prior to discharge or subjected to aerobic polishing, NPK removal, etc. by the industry prior to discharge to the receiving waterbody.

5. Used mainly at full-scale for treatment of wastewaters from the food and drinks sector. Growing recent application for more recalcitrant wastewaters.

6. EXAMPLE OF FULL-SCALE ANAEROBIC DIGESTER FOR INDUSTRIAL WASTEWATER TREATMENT ADM citric acid production plant in Co. Cork, Ireland. Wastewater characteristics:- 7000 m 3 /day 12000 mg COD/l 4000 mg sulphate/l

7. Digester specification:- Upflow, fully-packed anaerobic filter random-packed, polypropylene cascade rings; 7300 m 3 volume Diameter of 36 m, height of 12.4

8. Operational performance:- HRT of approximately 1 day 52% COD removal 81% BOD removal 30 m 3 biogas/day (66% CH 4 ) (corresponds to 18 l/min) Biogas is used for steam generation and space heating

9. North Kerry Milk Processing Plant in Co. Kerry, Ireland Wastewater characteristics:- 4000 m 3 /day 5000 mg COD/litre Digester specification:- Downflow, random-packed anaerobic filter, polypropylene rings 4500 m 3 volume

10. Operational performance HRT of approximately 1 day c. 90% COD/BOD removal Biogas used for electricity generation (combined heat and power plant).

11. Post treatment (activated sludge) prior to discharge Operated on a seasonal basis (March - October)

12. Granulation - a unique example of biofilm formation The UASB reactor was the first reactor design to employ granular anaerobic sludge Immobilisation of the bacteria is essential for reactor operation and ww treatment Intensively studied - yet incompletely understood

13. Although the UASB and related systems have been intensively studied for almost 2 decades, reactor instability due to biomass loss still causes problems - a common cause of reactor instability is poor granulation Granules are not formed/retained and biomass loss is not compensated by new growth; Biomass is lost by floatation Often biomass is not present as granules, but as poor settling flocs with low volumetric activity

14. What are granules? Granules consist of organised aggregates of microbial cells which form a complete methanogenic unit A granule contains all the microbial trophic groups necessary for complete breakdown of substrates to CH 4 Dense, well-settling aggregates

15. Granule formation occurs spontaneously when the appropriate conditions exist The exact requirements are still unclear Many factors implicated in granulation including: divalent metals, FeS, carbohydrates, hydraulic regime, ammonium, bacterial appendages, gas loading rate, surface tension of the liquid and extracellular polysaccharides

16. Will look in summary at current model for granule formation and some of the factors that influence a successful outcome

17. The microbial architecture of granules The bacteria involved in granule formation do not aggregate at random With microscopy and micro-electrode measurements, it has been established that granules have an organised distribution of bacterial species and activities

18. ANAEROBIC DIGESTION COMPLEX ORGANIC MOLECULES (e. g. polysaccharides, proteins) MONOMERS (e. g. glucose, amino acids) ORGANIC ACIDS, ALCOHOLS, KETONES ACETATE, CO 2, H 2 METHANE (CH 4 ) Hydrolytic/Fermentative bacteria Fermentative/Acidogenic bacteria Acetogenic bacteria Methanogenic bacteria

19. Fermentative or acidogenic bacteria are located on the outer edge of the granule (200µm) with the syntrophs and methanogens located in the centre The syntrophic acetogens must be located within 1-2 µm of methanogens in order to facilitate interspecies hydrogen transfer

21. How and why are Granules formed? Granule formation is complex - can divide the influencing factors into microbiological and environmental Microbiological factors include: EPS Kinetic parameters Cell surface characteristics Trace elements and nutrients Seed sludge

22. 1. The role of EPS in granule formation The importance of extra-cellular polymeric substances as a cementing substance is unchallenged - crucial in both the structure and function of biofilms and granules EPS is present in biofilms and aggregates and is present in a particularly structured sense in granular aggregates It cements cells together, and onto a substratum carrier

23. Besides its role in the structural integrity of the granules or biofilms it seems to function as a protecting agent against biocides such as the toxic chemicals which may arise in IWW In non-granular biofilms it protects against antibodies, antibiotics, prevent grazing and mediates initial cell attachment to surfaces Also acts as a storage material in times of starvation

25. There is a reported link between the presence of carbohydrates in the wastewater or feed and the maintenance of stable granular sludge These high-energy substrates will encourage the production of EPS In granules, EPS consists of polymeric carbohydrates, with a large amount of proteins, some lipid and DNA and some other biopolymers

26. EPS can be viewed as a highly hydrated gel, which is negatively charged at neutral pH This means that it effectively acts as an ion exchanger, strongly binding metal cations e.g. Ca 2+, and repelling anions This binding capacity could greatly reduce the diffusion of substrates through the granule, but only transiently - sites will become saturated

27. EPS and Floatation In practical terms for IWW AD the role of EPS may be vital, as acidogens on the outer layer produce the EPS This gives the granule a hydrophilic outer layer around an inner, hydrophobic layer of syntrophs and methanogens This hydrophilic layer helps to prevent gas attachment and floatation as well as cementing the granule together

28. 2. The role of kinetic parameters in granule formation Growth rates, substrate affinities etc. The granule represents a balanced ecosystem, this is not a straightforward outcome kinetically The growth rates of fermentative organisms are 5-10 times faster than syntrophs and methanogens - so environmental factors will greatly affect the likelihood of a stable community developing - delicate balance

29. The cell yields of methanogens and syntrophs are thus rate limiting for the growth of granules - loading rates and other parameters are based on this However, probably the most important kinetic determinant is the outcome of competition for acetate between two methanogens - Methanosaeta (ex. Methanothrix) sp. and Methanosarcina sp.

30. Acetate Competition and Granulation Methane can be produced via acetate or via H 2 /CO 2 - 70-90% of the biogas produced by anaerobic digesters is via acetate - the key CH 4 source Surprisingly however only two methanogens have ever been discovered that can utilise acetate - Methanosaeta and Methanosarcina

31. Methanosaeta concilli and M. Soehgennii are rod- shaped organisms which tend to grow as filaments Methanosarcina barkeri is coccoid and grows 3-6 times faster than Methanosaeta on acetate, but Methanosaeta is the dominant methanogen in granular sludge (up to 90% of methanogenic cells) Methanosaeta has a much higher substrate affinity and in a balanced anaerobic digester the concentration of acetate will be very low

33. The presence of Methanosaeta is believed to be crucial as the filaments provide a nucleus for granule initiation The role of cell surface characteristics in granule formation Hydrophobicity, electrophoretic mobility, and isoelectric point are used to predict the adhesion of bacteria to surfaces Hydrophobicity measured by water contact angle is the most commonly used - allows estimation of the surface energy of bacteria