International Journal of Applied Environmental Sciences

ISSN 0973-6077 Volume 12, Number 8 (2017), pp. 1561-1573

Research India Publications
http://www.ripublication.com

Influence of polymer addition on granulation in

upflow anaerobic sludge blanket reactor: A review

Vidya Singh, S. S. Narvi* and N. D. Pandey

Department of Chemistry, Motilal Nehru National Institute of Technology, Allahabad
211004, India.

*
Department of Chemistry, Motilal Nehru National Institute of Technology, Allahabad
211004, India.

Corresponding Author (S.S.Narvi)

Abstract
The upflow anaerobic sludge blanket (UASB) reactor has been used
successfully treat a variety of wastewaters. The aim of this review is to
synthesize and analyze information on how the process of granulation is
affected by environmental and operational conditions in the reactor. The
factors reviewed are strength of substrate, nutrients, multivalent cations and
heavy metals, exocellular polymer, and addition of natural and synthetic
polymers. Nature and strength of substrate in conjunction with intra-granular
diffusion largely determines the microstructure of the granules. The addition
of external additives as natural cationic polymers Chitosan as well as cationic
fraction of Reetha (Sapindus trifoliata) extract, synthetic polymers "AA 184
H" may enhance granulation in the upflow anaerobic sludge blanket reactors.
Keywords: Cations, Chitosan, Granulation, Polymers, UASB reactor.

INTRODUCTION
The performance of the upflow anaerobic sludge bed (UASB) system is highly
dependent on the granulation process with particular organic wastewater. The UASB
1562 Vidya Singh, S.S. Narvi and N.D. Pandey

process is attractive because of its compactness, low operational cost, low sludge
production, and production of methane. The application of UASB reactors for the
treatment of high-strength industrial wastewater containing easily hydrolyzed
substrates such as sugar industry wastes, distillery wastes, and brewery wastes had
been successful1-4. Performances of UASB reactors treating difficult-to-hydrolyze and
complex substrates such as phenols, effluents from food and milk processing plants,
gelatine manufacturing plants, and animal slaughterhouses were less than
satisfactory5-7. UASB process for the treatment of domestic wastewater (low strength)
also suffers from a number of shortcomings, such as long start-up time, poor gas
production, susceptibility to shock loading, inability to form self-immobilized
bacterial granules, and necessity for post-treatment of the effluent8-9. The granulation
process is commonly believed to be sensitive to the sudden change of environmental
and operational conditions10. Factors governing granulation have been extensively
studied on a variety of wastewaters. Some of these factors are operating conditions11-
13
, pH and alkalinity14-15, temperature16-17, strength and composition of wastewater18-
20
, reactor hydrodynamics17,21, presence of metal ions and trace metals22-25, presence
of polymers26-29, microbial ecology30-31, and production of exo-cellular polymeric
substances by anaerobic bacteria32-33.mA number of reviews are available on
functioning of UASB reactor34-35, its applicability to sewage treatment9,36, treatment
of some of the toxic wastes37, nutrient requirements for UASB process15 and
mechanism of granulation38-40. The main purpose of this review is to synthesize and
take a critical look at the effect of different environmental factors on enhancement of
granulation. The divalent cations such as calcium and iron may enhance granulation
by ionic bridging and linking exo-cellular polymers. However, their presence in
excess may lead to cementation due to precipitation leading to increased ash content
and mass transfer limitation. The addition of external additives such as ionic
polymers may enhance granulation in the upflow anaerobic sludge blanket reactors.

2. THE UASB TECHNOLOGY

2.1. The UASB reactor
The UASB reactor is initially with inoculums such as digested, anaerobic, granular
and activated sludge. Sludge enters from the bottom of the reactor. Under appropriate
conditions, light and dispersed Particles will be washed out while heavier components
will retain, thus minimizing the growth of finely dispersed sludge whilst forming
granules or flocs consisting of the inert organic, inorganic matters and small bacterial
aggregates in the seed sludge38. The schematic diagram of a UASB reactor is shown
in Figure 1.
Influence of polymer addition on granulation in upflow anaerobic sludge 1563

Fig. 1 Schematic diagram of a UASB reactor

The reactor consists of two parts: rectangular and cylindrical Column and a gas liquid
solid (GLS) separator. After a certain period (usually 2-8 months), depending on the
operating conditions and the characteristics of the wastewater and seed sludge a very
dense sludge bed which may be granular or flocculent in nature with high settling
properties develops. Above the dense sludge bed, there is a sludge blanket zone with
a much-diffused growth and lower particle setting velocities41. The biological
reactions take place throughout the highly active sludge bed and blanket zone. As the
flow passes upward, the soluble organic compounds in the influent are converted to
biogas consisting of mainly methane and carbon dioxide. The produced biogas and
the sludge buoyed by the entrapped gas bubbles are then separated from the effluent
by the immersed GLS separator, in which the baffles prevent as efficiently as possible
the washout of the viable bacterial matter or floating granular sludge by sliding the
settled solids back to the reaction zone38,42.

3. GRANULATION
The effectiveness and stability of a UASB reactor depends strongly on the initial
start-up, which in turn is main affected by numerous physical, chemical and
biological parameters19, such as the type of wastewaters, the operating conditions and
the characteristics, availability and growth of active microbial populations in the seed
sludge or inoculum. An acclimatization period is required before the full design
1564 Vidya Singh, S.S. Narvi and N.D. Pandey

organic loading rates can be applied to inoculate the seed sludge to the operating
conditions. This period which is typically 2-8 months2 is rather long and has been the
major hitch of the industrial applications of UASB reactors.

4. ENHANCING FACTORS OF GRANULATION

4.1 Substrate characteristics
No layer in the granules was reported to form with propionate, peptone, ethanol, and
glutamate as substrates6,18,43. The granules also do not show any layered structure in
the presence of an inhibitory substrate such as phenol or lindane43. Methanogenesis is
reported to be the rate-limiting step in the anaerobic degradation of the non-inhibitory
substrate whereas with the inhibitory substrate, acetogenesis, is identified as the rate-
limiting step11,44. Low substrate concentration in the feed may result in substrate
limitation at the core of the larger granules as majority of the substrate is utilized near
the surface40.

4.2 Multivalent cations and heavy metals

The cations may accelerate this process through bridging between negatively charged
groups on cell surfaces and linking exocellular polymers38,45,46. In addition,
multivalent cations condense the diffused double layers and facilitate flocculation due
to Vander Waals forces46. The predominant binding groups for metals on the surface
of bacteria are carboxyl and amino groups in proteins.

4.3 Inoculums
In order to select the best inoculums source for a specific type of wastewater, the
toxicity and biodegradability tests as reported in the literature can be used24. Although
a UASB reactor can perform efficiently without granules, granule formation during
start-up lends a decided advantage for its ability to provide high COD removal
efficiency within a much shorter period, allowing treatment for larger volumes of
wastewater. The importance of granules in the operation of UASB reactors has led to
an increasing number of studies on the relevant theories and mechanisms of anaerobic
granulation38,46. According to46 the initial development of granules can be divided
into four steps: (1) Transport of cells to a substratum (i.e. an uncolonized inert
material or other cells); (2) Initial reversible adsorption to the substratum by
physicochemical forces; (3) Irreversible adhesion of cells to the substratum by
microbial appendages or polymers and (4) Multiplication of the cells and
development of granules.
Influence of polymer addition on granulation in upflow anaerobic sludge 1565

4.4 Different types of cations

Presence of divalent and trivalent cations exert positive impact on granulation process
by neutralizing negative charges on bacterial surfaces, serving as cationic bridges
between bacteria. Investigators24,47successively studied the roles of a few specific
multivalent cations on granulation using the same UASB reactor of 7.3 L in volume
for treating synthetic wastewater with influent COD concentration of 4000 mg/L.
They found that with an optimum concentration of iron (Fe2+) at 300 and 450 mg/L,
the granulation process was accelerated; higher Fe2+ concentration in the feed (more
than 450 mg/L) resulted in excessive deposition of minerals on the granules, therefore
deteriorating the bacterial specific activity24. As with the addition of calcium (Ca2+),
the optimum concentration of Ca2+ at 150-300 mg/L enhanced biomass accumulation
and granulation process by accelerating the adsorption, adhesion and multiplication of
the granulation process47. The addition of aluminium (Al3+) at 300 mg/L resulted in
the formation of large granules and shortened the granulation time by approximately
one month25,48 , examined the effect of adding aluminium chloride on the treatment of
low-strength (650-750 mg/L) synthetic wastewater. It is found that the addition of
Al3+ at 200-300 mg/L (either continuously throughout the operation or during the
start-up) adversely affected COD removal efficiency and growth of agglomerates. A
smaller concentration of Al3+ at 50 mg/L, did not affect the COD removal efficiency
but adversely affected the growth of agglomerates48. These studies25,48 have critically
suggested that the optimum concentration of multivalent cations is dependent on the
influent COD concentration. Consequently, more research studies are needed to
determine their relationships and roles in the granulation process. Magnesium (Mg2+)
ion is also influencing the properties of denitrifying sludge and granulation. Two
upflow anaerobic sludge bed (UASB) reactors were used, R1 was continuously
supplied with 50 mg L1 of Mg2+ in the wastewater and R0 was used as a control
without magnesium. On the 258th day, the sludge properties in R 0 and R1 were
analyzed when the nitrogen loading rates were both 36.0 kgN m 3 d1 and the
nitrogen removal rates of R0 and R1 were 33.8 and 34.6 kgN m3 d1, respectively.
The diameters of R0 and R1 were 0.95 and 1.35 mm, settling velocity (74.5 20.8
and102.0 22.6 mh1), extracellular polymeric substance (60.8 2.1 and
76.0 2.7 mg g1VSS) and specific denitrifying activity (794.4 9.6 and 825.6 4.9
mgN d1) were also compared. These results addressed that the addition of
magnesium to denitrifying reactor is an effective alternative to enhance sludge
properties49.

4.5 Different Polymers

In addition to the use of multivalent cations, the natural polymers, such as water
extract of the Moringa Oleifer seeds (WEMOS)26, Chitosan29,50, Reetha extract29 and
1566 Vidya Singh, S.S. Narvi and N.D. Pandey

powdered bamboo-charcoal51 as well as commercial and synthetic polymers, such as

the commercial cationic polymer AA184H27-28 and organic-inorganic hybrid
polymers52 also showed promising results in enhancing the start-up and granulation in
UASB reactors. A few cationic polymers reported to enhance granulation are natural
cationic polymers Chitosan29,53 and cationic fraction of Reetha (Sapindus trifoliata)
extract29, synthetic polymers AA 184 H and Percol 76353. The efficiency of Chitosan
was found to be better than the synthetic polymer Percol 76353 and natural polymer
Reetha extract29.

4.5.1 Moringa Oleifer seeds (WEMOS)

The enhancement of start-up of self-inoculation by adding a water extract of Moringa
oleifera seeds (WEMOS) by26. M. oleifera is a tropical plant belonging to the family
Moringaceae. WEMOS is effective in flocculating organic matter and removing
microorganisms from raw water54. It is currently used to treat drinking water, where it
has shown its efficiency as a coagulating agent. The active agents of WEMOS are
dimeric cationic proteins with a molecular weight of 13 kDa and an isoelectric point
between 10 and 11. The WEMOS solution contains several types of carbon sources,
e.g. carbohydrates, lipids and lignin, and also nutrients such as N, P and metal ions
which are capable of supporting microbial growth. It was therefore, hypothesised that
a continuous supply of WEMOS to a UASB reactor treating raw domestic wastewater
would result in an enhancement of its start-up period. Water extract of Moringa
oleifera seeds (WEMOS) was used to enhance the start-up of a self inoculated upflow
anaerobic sludge blanket (UASB) reactor treating raw domestic wastewater. Two
reactors labelled control (RC) and WEMOS addition (RM) were started without
special inoculum. Both reactors were fed continuously for 22 weeks with domestic
wastewater containing an average total chemical oxygen demand (COD) of 320 mg
O2/L and suspended solid (SS) of 165 mg/L. The reactors operated during the entire
experimental period at 29C and at a hydraulic retention time (HRT) of 4 h. The RM
reactor received 2 ml WEMOS per litre of influent. WEMOS solution was prepared
on the basis of 2.5% (w/v) ground M. oleifera seeds in water. The results of 22
weeks operation showed an improvement in the performance of the RM compared to
that of the RC. The dosage of WEMOS in the feed (1) shortened the biological start-
up period by 20% (2) increased acidogenic and methanogenic activity by a factor of
2.4 and 2.2 respectively (3) increased the specific biogas production by a factor of
1.6,(4) favoured fast growth of the sludge bed and (5) allowed the aggregation of
coccoid bacteria and growth of microbial nuclei, which are precursors of anaerobic
granulation.
Influence of polymer addition on granulation in upflow anaerobic sludge 1567

4.5.2 Chitosan and Reetha extract (bulk, cationic fraction, and anionic fraction)
Chitosan is a modified polysaccharide, mostly produced by alkaline deacetylation of
chitin. In a UASB reactor, granules start to form with the aggregation of acidogens,
and with methanogens enclosed inside, leading to the formation of an elastic
hydrophilic layer and a hydrophobic inner core, thus decreasing the washout of
methanogens.
This was attributed to the polysaccharide structure of the Chitosan, similar to
exocellular polymer found in aggregating anaerobic sludge and the water-absorbing
nature of the polymer55. The effectiveness of some water-absorbing polymers in the
enhancement of granulation56. Due to this nature, Chitosan has shown positive results
in enhancing sludge granulation and has exceeded Percol 763, as well as both the
cationic and anionic fraction of Reetha extract in treating synthetic wastewater29. In
the latter, large granules were formed and their stabilities were shown to be stable,
indicating the low possibility of substrate-diffusion limitations with low organic
loading. The effects of chitosan characteristics (i.e., degree of deacetylation [DD] and
molecular weight) and environmental conditions (i.e., ionic strength and pH) on the
flocculation of anaerobic sludge were investigated. The results showed that chitosan
enhanced the flocculation of sludge, and the flocculation efficiency depended on both
the degree of deacetylation and molecular weight. Chitosan with 85% DD was more
effective than that with 70% DD, as the former required a lower dose to obtain 90%
flocculation at all studied pH values. In addition, low molecular weight chitosan
enhanced the flocculation better than high molecular weight chitosan. The increase in
ionic strength (up to 0.1 M) of the suspension helped reduce destabilization that
occurred when chitosan was overdosed. In general, chitosan has potential to be used
as an effective cationic bioflocculant, which is able to function either in acidic or
neutral conditions, and very small amounts of chitosan (less than 4 mg/g dried sludge)
are required50. Concomitant early granulation with chitosan addition under a
syntroph-specific substrate and enhancement of extracellular polymeric substances
(EPS) production were aimed to build anaerobic granules with high syntrophic
activities in a short period. Two laboratory-scale upflow anaerobic sludge blanket
reactors were operated as control (R1) and chitosan addition (R2) reactors during
early granulation (phase 1). Chitosan decreased the negativity of microbial surface
charges (zeta potential) to 10.5 mV on day 58 which led to increases in average
diameter sizes, nuclei and granule ratio of approximately 115 m, 55.1% and 8.2%,
respectively. While zeta potential in R1 slightly changed, this resulted in less
microbial aggregation. Although microbial aggregation in R2 was rapidly triggered
by chitosan addition during phase 1, its structure was clumpy with rough surface due
to lack of EPS. Substrate switching to glucose increased polysaccharides EPS during
phase 2 which was synergistically improved on the structural characteristics of
microbial aggregate in R2, that is, more spherical and compact, with a smoother
1568 Vidya Singh, S.S. Narvi and N.D. Pandey

surface. Rapid-growth microorganism was also boosted, which then dominated the
outer layer of the aggregate. The Archaea clumps were observed at a deeper layer and
were surrounded by Eubacteria, presumably acetogens, indicating a syntrophic
relationship due to substrate association between these microbial groups57.

4.5.3 Commercial cationic polymer AA184H

The impact of a commercial cationic polymer AA184H at concentrations 5-20
mg/L on granulation and organics removal efficiency in treating synthetic sludge with
COD 5000 mg/L with dosage of 20 mg/L27-28. The organic loading capacity of the
UASB reactor was increased from 19.2 to 25.6 g COD/L/d28 while with dosage of 80
mg/L, the start-up period was shortened and the granules in UASB reactors were
strengthened and exhibited the best settleability at all studied organic loading rates
(OLRs) (2-40 g COD/L/d leading to increased organic removal efficiency and loading
capacity of the UASB system27.

4.5.4 Organic-inorganic hybrid polymers

From Synthesized organic-inorganic hybrid polymers and added to the UASB
reactors52. It was found that granular sludge was formed within 5 min. The granules
were stable throughout the operation and the COD removal efficiency was as high as
90% at even up to 18 g COD/L/d of organic loading rate.

CONCLUSIONS
The optimum range of organic loading rate (OLR) and hydraulic retention time
(HRT) can only be decided after considering the influent characteristics and other
operating conditions. Layer formation in granules is due to interaction of
intragranular diffusion and degradation kinetics of the substrate. Thus, the granule
microstructure is governed by the substrate type and strength. The presence of
adequate nutrients and metals is essential for granulation. Calcium and iron may
enhance granulation but are also capable of causing mass transfer limitation when
present in large quantities. External addition of ionic natural and synthetic polymers
can be used to enhance granulation. They help in the initial stages of formation of
granules. Another challenge is to shorten the start-up time of the reactor by enhancing
granule formation. Various external additives have shown promising results in this
direction, however, most of these studies are limited to laboratory-scale reactor. The
effect of these additives should be investigated in pilot-scale reactors along with the
economics of the additives.
Influence of polymer addition on granulation in upflow anaerobic sludge 1569

ACKNOWLEDGMENTS
Authors are grateful to MHRD, New Delhi MNNIT, Allahabad for their help in
compiling and preparing this manuscript.

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