- Van Den Hende et al.

2011: experiment with flue gas

o Data matching to outdoor high rate algal ponds and valorisation
of the biomass has yet to be investigated, before the stage is set
for the industrial implementation of MaB-floc ponds.
o Further research is needed to clarify whether flue gas sparging
inhibits bacterial ammonium oxidation, enhances microalgal
nitrate uptake and/or decreases the ammonium inhibition of
nitrate uptake in light conditions.
o Bioflocculation can lower the biomass harvesting costs
by providing a nearly cost-free first harvesting step, as only (a
short) time is the prerequisite. The need and choice of a second
harvesting step, such as centrifugation, will depend on the
application and composition of the biomass.
o More accurate methods for the determination of microalgae and
bacteria are needed, especially in mixed and flocculating
biomass samples.
- Van Den Hende et al. 2014: industrial w.w. in sequencing batch
reactors.
o Phosphorous was limiting the nitrogen removal in manure
treatment wastewater. The addition of dark-coloured CAS
influent to light-coloured CAS effluent of the manure treatment
could enhance the TN:TP ratio, but will also lead to a darker
wastewater colour, thereby affecting autotrophic microalgae
growth. A balanced mixture for manure treatment wastewaters
needs to be determined.
o Further research is needed to confirm whether this was because
UASB sludge particles were removed during settling in the
SBR but not oxidised during BOD5 determination, and/or
photosynthesis-induced metabolism occurred which resulted in
some surplus TCOD that was vulnerable to biologic
degradation.
o On average more TN was removed compared to TC, as can be
seen from the low TC removal rate: TN removal rate ratio.
Further research is needed to confirm whether this is due to

ammonia volatilisation at a pH level of above 9 or

denitrification during anoxic/anaerobic dark periods.
o It should be verified if discharge norms can be met for
the nutrient-rich wastewater of the food-processing industry by
increasing the HRT.
- Van Den Hende et al. 2014: up-scaling
o This means that for each mol TN removed,
1.52 1.43 mol TC and 2.38 1.33 mol TOC was removed.
The latter values are low compared to the TC:TN ratio of
micro-organisms of approximately 6 (Geider and La Roche,
2002). This suggests that the N removal could not be due to
biomass growth only. More research is needed to confirm
whether denitrification in anoxic microniches of the flocs, as in
activated sludge was an additional mechanism for N removal.
o This means that not all C and N removal could have been due to
biomass growth. More research is needed to confirm which of
the following removal mechanisms play a role in this: TN
removal via denitrification, VSS removal by predators, TC
removal via CO2 emission to air and CaCO3 precipitation.
o The nitrate concentration of the effluent (55.0 8.60 mg N
NO3 L1) was largely above the current discharge norm of 33.9
mg NNO3 L1. The question remains whether this was due to
nitrification, excretions by MaB-floc predators and/or lysis of
organic compounds catalyzed by enzymes excreted by
cyanobacteria, such as protease and urease.
o The supersaturated DO values up to 24 mg L1 (Fig. 5)
demonstrate photosynthetic aeration by MaBflocs and BOD5
underloading of 12M. These supersatured DO values can be
toxic for microalgae (Richmond, 2004). Therefore, during
further optimisation research should investigate the potential of
increased BOD5 loadings on the reactor performance, for
example by altering the influent pre-treatment step.
o Negative NO2 REs were obtained, despite the aerobic
conditions needed for nitrification. Further optimization to
lower the NO2 concentrations is crucial. Since the NO2

concentration in the indoor fish tanks was lower than 1 mg L1,

the NO2 must have increased during storage in the influent
buffer tanks. Therefore a possible strategy to enhance the NO2
of the effluent would be to decrease the HRT of the influent
buffer tanks by discontinuous feeding of the MaB-floc SBR
during the day.
o Conclusion:
Firstly, up-scaling to outdoor conditions strongly
increased the effluent pH. Outdoors, flue gas sparging
was needed to obtain an effluent pH below the discharge
norm. Despite being advantageous in terms of
greenhouse gas mitigation, flue gas sparging in
wastewater treatment raceways represents an extra cost
(up to 20% of the capital expenditures; Zamalloa et al.,
2011 ). In certain countries, such as Belgium, this cost
cannot be compensated by revenues for CO2 credits
because no such credits can be obtained for flue gas
scrubbing in open ponds.
Third, up-scaling increased the HRT to 4 days. This
means that per m3 indoor fish tank, a microalgae
raceway pond area of 1 m2 would be needed (daily
discharge of 10% of the fish tank water). To overcome
potential problems of regarding shortage of availability
of land, raceway ponds on the roof of the indoor
aquaculture facility could be a worthwhile option to
investigate. If year round wastewater treatment is
targeted, pond heating with waste heat will be needed.
o Microscopic observations showed that a large amount
of crystals were present in the MaB-flocs (Fig. A1). Further
research is needed to confirm the nature of these crystals.
o Future research should focus on nitrogen removal and biomass
valorisation.
- Sutherland et al. 2015: enhancing microalgae
o Organic nutrient loading rate, hydraulic retention time, pond
depth, CO2 supply, as well as vertical and horizontal mixing

velocities are the main operational parameters that can be

modified in a HRAP. In particular, given that reported
photosynthetic efficiency is, at best, only 1/5 of the theoretical
maximum, further research is necessary to understand how the
operational parameters impact on the physiology,
photosynthetic capability and productivity of HRAP
microalgae.
Mixing frequency, as well as the mixing velocity, is
likely to play a critical role for maintaining desirable
large colonies in HRAPs. Understanding how the
frequency of mixing events affects the performance of
microalgae, including photosynthesis, productivity,
nutrient removal efficiency, as well as physiological and
morphological adaptations, is important for enhancing
wastewater treatment and biomass yields.
Under medium frequency light fluctuations (ranging
from seconds to minutes), such as those expected in fullscale wastewater HRAPs, the effects of enhanced vertical
mixing on photosynthesis and productivity have been less
conclusive.
While the effects of HRT and nutrient loads have been
investigated with respect to nutrient removal and biomass
production, their effects on microalgal physiology have
not been investigated, to date. Increased HRT would lead
to increased biomass concentration and therefore reduced
pond light climate. This may negatively impact on light
absorption, photosynthesis and ultimately biomass
production as microalgae become low light adapted.
o Available land is often limited in most countries and operating
sufficient numbers of HRAPs at the shallowest pond depth may
not be economically, or socially, viable. In order to reduce their
footprint, wastewater HRAP systems have been designed to
retrofit into existing facultative pond systems. There is a need
to understand the performance of HRAPs with respect to
optimizing microalgal photosynthesis, nutrient removal

efficiency and areal productivity and the trade-offs with

increasing pond depth.
o While there have been some mesocosm and pilot-scale studies
evaluating ways to increase both light and carbon in wastewater
HRAPs (as discussed above), there has been a lack of research
in this regard at full-scale, to date. Research at full-scale on
both improved light and carbon availability should be the main
focus of future studies. Full-scale research would help to
answer the question of whether implementing operational
modifications can be done cost-effectively. CO2 addition and
enhanced mixing, while identified as beneficial to microalgal
productivity at pilot-scale, could substantially increase capital
and operational costs and the benefits, in terms of biomass
yield, may not out way the costs unless improvements in these
technologies are made.
- Su et al. 2012: algae/bacteria ratios.
o Some algae-based bio-systems were used to treat highly
concentrated municipal wastewater and the corresponding COD
was removed from initial 2500 mg/l to around 250 mg/l (Zhou
et al., 2011; Li et al., 2011). The performance of the algae
sludge system proposed in this study for high concentration
wastewater treatment could be our further research goal.
- Park et al. 2011: wastewater treatment HRAPs for biofuel
o Since wastewater treatment HRAP systems are already an
economically viable technology for efficient tertiary-level
wastewater treatment, they could potentially be the testing
ground to develope large-scale algal production, harvest and
biofuel conversion technologies that will be latter used when
higher fossil fuel costs make purpose built algal biofuel
production systems economical. Therefore, both fundamental
and field-scale research is urgently needed to optimise algal
production and harvest from wastewater treatment HRAPs,
however for these systems this must be achieved while
maintaining high effluent water quality.

o Further research is required in large-scale wastewater treatment

HRAPs using cheap/free CO2 sources (flue gas or biogas) to
minimize operational costs.
o The influence of zooplankton grazing and parasitism (fungal
and viral) on HRAP wastewater treatment and algal
production requires further study. A greater understanding of
how these organisms interact with HRAP algae may lead to the
development of effective control methods.
o The physiological properties of colonial algae and exact
mechanisms of the aggregation have yet to be determined.
- Garcia et al. 2000: HRAP for nitrogen removal.
o From an engineering point of view, the optimal size of a HRAP
for nitrogen removal requires a mathematical modeling that
correlates effluent TN with influent TN, through environmental
conditions and HRT. The design models normally available are
based on BOD5 loading; a further refinement should be made
to include nitrogen removal processes, as indicated in this
study.