south african journal of chemical engineering 24 (2017) 62e70

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South African Journal of Chemical Engineering

journal homepage: http://www.journals.elsevier.com/
south-african-journal-of-chemical-engineering

Bioremediation of acid mine drainage using algae

strains: A review

J.K. Bwapwa*, A.T. Jaiyeola, R. Chetty

Mangosuthu University of Technology, Engineering Faculty, P.O Box 12363, Jacobs 4026, Umlazi, Durban,
South Africa

abstract

Acid mine drainage (AMD) causes massive environmental concerns worldwide. It is highly acidic and contains
high levels of heavy metals causing environmental damage. Conventional treatment methods may not be effective
for AMD. The need for environmental remediation requires cost effective technologies for efficient removal of
heavy metals. In this study, algae based systems were reviewed and analyzed to point out the potentials and gaps
for future studies. Algae strains such as Spirulina sp., Chlorella, Scenedesmus, Cladophora, Oscillatoria, Anabaena,
Phaeodactylum tricornutum have showed the capacity to remove a considerable volume of heavy metals from AMD.
They act as hyper-accumulators and hyper-adsorbents with a high selectivity for different elements. In
addition, they generate high alkalinity which is essential for precipitation of heavy metals during treatment.
However, algae based methods of abating AMD are not the ultimate solution to the problem and there is room for
more studies.
2017 Mangosuthu University of Technology. Published by Elsevier B.V. on behalf of Institution of Chemical
Engineers. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-
nc-nd/4.0/).

Keywords: Acid mine drainage; Algae strains; Contamination; Heavy metals; Bioremediation

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
2. Review of acid mine drainage: summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
3. Absorbent and adsorbent properties of algae biomass . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
4. Previous studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
5. Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66
6. Shortcomings of phycoremediating using aquatic plants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
7. Unsolved issues and novel approach . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
8. Challenges and future prospective . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68
9. Conclusions and recommendations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68

* Corresponding author.
E-mail addresses: joseph@mut.ac.za, josephkapuku@gmail.com (J.K. Bwapwa).
Received 12 July 2016; Received in revised form 27 April 2017; Accepted 25 June 2017
http://dx.doi.org/10.1016/j.sajce.2017.06.005
1026-9185/ 2017 Mangosuthu University of Technology. Published by Elsevier B.V. on behalf of Institution of Chemical Engineers. This is
an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
 south african journal of chemical engineering 24 (2017) 62e70
1. Introduction
Acidic solutions can be formed by natural decomposition of
sulfide materials through oxidative processes in effluent
wastewater. The leaching of heavy metal contaminants from
within a rock matrix can be a result of high acidity levels of
effluent wastewater. These processes can be catalyzed by
contact with pre-existing environmental contaminants. These
contaminants in very small quantities form activated com-
plexes with various heavy metal atoms encouraging their
ionization, dissolution and suspension in the liquid waste
medium. These effluents can be released into the environment
by spillage, overflow, percolation and intentional discharge. It
has adverse effects on environmental health because of the
heavy metals pollution downstream from the source.
Regarding acid mine drainage (AMD), the high concentra-
tions of heavy metals cause major environmental concerns
characterized by contamination of various ecosystems due to
its leaching capacity and the presence of very active bacteria
making it self-perpetuating (Kalin et al., 2006). There is a ne-
cessity for AMD treatment to prevent disastrous conse-
quences in the environment. Therefore, effective methods of
trapping and sequestering heavy metals from effluent to
prevent downstream contamination and remediate the local
environment are required. Various bioremediation methods
based on the ability of plants to take up and accumulate heavy
metals were suggested. There are two distinct strategies to
remediate the issues regarding AMD treatment. The first is the
conventional treatment process in which the effluent is
collected then biologically and chemically treated in a
centralized wastewater treatment plant. The second involves
the channeling of the effluent through natural or constructed
wetlands in which microbes or cells such as anaerobic and
aerobic microorganisms and various algae strains can be used
to treat the wastewater passively. In this study the focus will
be on algae based treatment for AMD. An effective treatment
for AMD should be self-renewing; the use of algae based
treatment is ideal because of its sustainability.
Algae as decontaminating agents offer several advantages
including low costs, easy manipulation, non-polluting, rela-
tively simple recovery of the metal contaminants for recy-
cling, and are not a source of secondary waste (Kalin et al.,
2006). Conceptually, the algae should grow in the contami-
nated effluent then the algal biomass and water should be
separated and dried to recover the concentrated metals con-
tent by conversion to oxides or other recoverable salts.
Alternatively, the recovered dry algal biomass can be stored
for future use or sequestered. Also, algae biomass can be
disposed in municipal waste landfills to reduce its environ-
mental footprint and boost the potential of producing biofuel
(Edmundson and Wilkie, 2013).
Phycoremediation is the process of employing macro or
microalgae for wastewater treatment. It has many advantages
over the conventional methods, which are very costly, energy
consuming and generating high amount of sludge hence it is
accepted throughout the world (Ghosh and Singh, 2005; Abdel-
Raouf et al., 2012). This method involves the use of macro or
microalgae for the removal or biotransformation of pollut-
ants, including nutrients and xenobiotics from wastewater
(Ahmad et al., 2013). Over the last few decades studies were
undertaken to apply microalgae such as Chlorella, Chlamydo-
monas, Spirulina, Scendesmus, Nostoc and Oscillatoria for waste-
water treatment (Dubey et al., 2011; Sharma and Khan, 2013).
63
The fundamental assumption is that the microalgae are
versatile to convert the contaminants into non-hazardous
resources, enabling the treated water to be recycled or
reused or safely discharged (Rao et al., 2011). This technology
is low in cost and it is an effective approach to remove excess
nutrients, contaminants in wastewater and generating
possibly useful biomass (Sengar et al., 2011). Therefore pre-
sent investigation focuses on analyzing various species that
are capable of removing contaminants from AMD or toxic
compounds from acid mine drainage. It also looks at the
mechanism of removing contaminants, the operating condi-
tions and the critical aspects of the technology. Also, to clearly
describe the effectiveness and potential of algae based tech-
nology for removal of contaminants in acid mine drainage.
Today algae based treatment is used in combination with
secondary treatment. Microorganisms work in symbiosis with
algae cells for an effective treatment. The study will
contribute also in emphasizing on the current role of algae in
wastewater treatment.
2. Review of acid mine drainage: summary
Although various natural environmental effects contribute to
the formation of acids in the environment, AMD as a result of
human activities can be attributed in large part to the oxida-
tive decomposition of exposed pyrite (iron sulfide, FeS2) by
water and oxygen as presented in equations (1)e(4):
FeS2 7=2O2 H2 O/Fe2 2SO2
4 2H (1)
Fe2 1=4O2 H /Fe3 1=2H2 O (2)
Fe3 3H2 O/FeOH3 3H (3)
FeS2 14Fe3 8H2 O/15Fe2 2SO2
4 16H (4)
The oxidative reaction converts the solid pyrite to dis-
solved ferrous iron ions (Fe2) and the equivalent of two units
of aqueous sulfuric acid (2H ions and 2SO2 4 sulfate ions
dissolved in the water). The ferrous ions, on exposure to dis-
solved or atmospheric oxygen, undergo further oxidation to
produce ferric ions (Fe3) which then react directly with pyrite
to increase the acid content of the water and the establish-
ment of acidophilic bacteria colonies further promotes the
acidification process (Costello, 2003).
Gaikwad and Gupta (2008) associate acid mine drainage,
also known as abandoned mine drainage, to the reaction of
water and oxygen specifically with coal mining. Water passing
through the rocks from mining operations that have been
deposited on the surface and the underground voids left
behind by mining activities causes the formation of acidic
drainage effluents. This occurs by the reaction involving ox-
ygen, water and pyrite sulfidic and non-sulfidic minerals to
produce acid sulfate rich wastewater know as AMD. Kuyucak
(2002) terms the AMD phenomenon as acid rock drainage
(ARD) and notes that the same chemical reactions can take
place in roads and bridge construction, and in the construc-
tion of tunnels through rock formations.
With the formation of net acidity, effluent water becomes
increasingly laden with Fe, Mn, Al, Zn, Cu, Ni, Pb, As and Cd as
the principle heavy metal contaminants. Ben and Baghour
(2013) found that Cd is typically the most mobile and there-
fore has the highest bioavailability. Furthermore, the presence
64 south african journal of chemical engineering 24 (2017) 62e70
of microorganisms promoting geochemical processes together
with mineral oxidation reduces the amount of dissolved oxy-
gen in water (Bhattacharya et al., 2006).
Generally, the most common uses of algae as phycor-
emediators reported in the literature are their character of
being alkalinity boosters rather than as direct absorbers. The
removal of heavy metal contaminants by algae species assists
in monitoring the levels of heavy metals in an environment.
However, in terms of efficiency and ease of maintenance, the
turnover of algae due to toxicity of the accumulated heavy
metals makes continuous processes much more attractive.
The performance of such systems over time is also more
easily monitored. According to Costello (2003) many examples
of these passive systems are operational and are still under-
going research to improve their functional characteristics.
The use of various algae strains to remove heavy metals
from AMD has only more recently become the subject of closer
scrutiny. Gok and Yel (2009) carried out studies of the kinetic
characteristics of heavy metals uptake by microalgae. These
characteristics would be of fundamental importance in
designing effective and long-lasting phycoremediation sys-
tems. Perales-Vela et al. (2006) completed a study on heavy
metals detoxification mechanisms in microalgae examining
the biomolecular way by which the algae isolate and sequester
those environmental materials to which they are exposed.
They particularly examined the formation of metallothionein
peptides in the algae.
3. Absorbent and adsorbent properties of
algae biomass
The removal of heavy metals and sulphates by algae is very
flexible. It depends on the metal type, the taxon and age of
material (Novis and Harding, 2007). Optimum heavy metals
and contaminants removal vary with season type (Elbaz-
Poulichet et al., 2000; Brake et al., 2004). Seasons of the year
can highly influence the contaminants removal because of
parameters such as light and temperatures. Algae are very
sensitive when it comes to parameters related to seasons such
temperature and light intensity.
The absorbency and adsorbency mechanisms are
commonly used by algae species to remove nutrients, heavy
metals (depending on the specie type) and other minerals
from wastewater. While these elements are removed from
wastewater algae growth takes place because species need
also nutrient elements to grow. Some are taken by the surface
and other taken into the inner cells as presented in Fig. 1.
According to Mehta and Gaur (2005), algae in aqueous solution
is very effective for the removal of low concentration of metal
Fig. 1 e Absorption and adsorption scheme mechanism
from a microalgae cell. Adapted from Kaplan (2013).
ions and also bio accumulate these metals within their cells
more specially in the cell vacuoles and/or in the intercellular
spaces (Afkar et al., 2010; Chen et al., 2012; Kumar and Gaur,
2011). Cladophora glomerata and Oedogonium rivulare are
amongst species used to continuously remove Co, Ni, Pb, Cd,
Mn, Fe, Cr and Cu from wastewater (Vymazal, 1984).
This ability of algae to remove heavy metals varies with the
different strains of algae and it is generally in the following
green algae descending order Chlorophyta e Phaeophyta (red
algae) Rhodophyta (Al-Shwafi and Rushdi, 2008). Furthermore,
it was found that the lifeless algae biomass adsorbs more
metals than living algae (Mehta and Gaur, 2005). Stigeoclonium
sp. a fresh water algae can survive in mining water with high
concentration of Zn of about 10 mM and it is also effective in its
removal (Pawlik-Skowronska, 2001).
In living algal cells, trace metals intracellular are accu-
mulated by active biological transport (Ajjabi and Chouba,
2009; Kiran and Thanasekaran, 2011). The presence of
various variety of algae coupled with their multilayer cell
walls make them suitable as a cheap source of adsorbent for
heavy metals (Bilal et al., 2013; Gupta et al., 2015; Wang and
Chen, 2009). Algae are among the most commonly found
photosynthetic eukaryotes (Anastopoulos and Kyzas, 2015)
that have the ability to survive in both fresh and marine
brackish waters.
Spirogyra algal species was found to have a removal effi-
ciency of 58e85% for Cu (II) at initial concentration of 20 mg/L
after 30 min (Bishnoi and Pant, 2004). Caulerpa lentillifera dried
green microalgae is also an effective bio adsorbent for multi-
ple metals removal in aqueous solutions (Pavasant et al.,
2006). Brown algae Turbinariaornate and green algae Ulothrix
zonata are also very effective adsorbents in the removal of
heavy metals (Nuhoglu et al., 2002; Vijayaraghavan and Prabu,
2006; Djati Utomo et al., 2016).
Regarding the removal of sulphates, it is reported that
microbial sulfate reduction generates alkalinity and the for-
mation of metal sulphides allows the precipitation of metals
from the solution. AMD environments are naturally carbon
limited (Koschorreck, 2008), thus the addition of suitable car-
bon sources is required to promote the activity of SRB. For SRB
to survive and for their better growth, nutrients are impera-
tively required. Furthermore, sulphates reduction and metal
precipitation have to take place to allow an effective growth
and survival. The expansion of technology using SRB for AMD
remediation is constrained by costs and obtainability of a
carbon source (Molwantwa et al., 2000).
Previously, biotechnologies have been used to recycle
wastewater with the help of microbial populations for the
removal of pollutants. In the case of AMD, the most studied
bioremediation for removal of heavy metals is based on
sulphates reducing bacteria (Kiran et al., 2017). In this pro-
cess heavy metals are removed through the production of
metal sulphide precipitates. This technology was successful
on a large scale for heavy metal removal at low pH values,
stable sludge, very low operating costs and minimal energy
consumption. Therefore, according to Sheoran et al. (2010),
SRB could be a very effective alternative treatment method
for AMD on sites with no power but are not suitable for sites
with low temperatures such those under extreme winter
conditions.
However, it has the weaknesses of long residence times
that can take weeks. Also, it requires a continuous supply of
organic substrates and the use of large steel bioreactors
(White et al., 1997; Sheoran et al., 2010). In addition, as it well
 south african journal of chemical engineering 24 (2017) 62e70
known for any anaerobic process, the start-up is time
consuming, requires an appropriate microbial population to
work effectively and capital costs can be high. Hoffmann
(1998) concluded that algae-based treatment can be used for
the removal of inorganic substances from effluents. However,
the greatest advantage of SRB is the simultaneous removal of
both metals and sulphates, which does not occur with the
traditional chemical processes (Kiran et al., 2017).
4. Previous studies
The pollution of the aquatic environment by the discharge of
AMD containing heavy metals has been a worldwide chal-
lenge and a concern for many decades (Nriagu and Pacyna,
1988). It was confirmed by many studies undertaken world-
wide on the effects of AMD on the environment. Due to a
number of negative effects produced by AMD on the envi-
ronment, many countries have developed policies and regu-
lations regarding the discharge of AMD. Through these
policies and regulations, standards are suggested in order to
prevent threat to human health and disastrous environmental
impact, dropping concentrations in effluents, and developing
cost effective technologies (Fan, 1996).
Generally, AMD treatment requires the use of lime/lime-
stone or any other alkaline agent in order to raise the pH and
allow the precipitation of metals as well as various metal
hydroxides. However, large amounts of sludge is generated
from this method. Therefore, it does not effectively solve the
problem of sulphates removal when compared to the perfor-
mance of SRB regarding the same issue.
Basically, the treatment of AMD is based on either sepa-
ration such as precipitation, adsorption and filtration or any
other separation or reaction process used for water and
wastewater treatment. The weakness of these conventional
processes includes low removal capacity, lack of selectivity
and intolerance to organic substances. Furthermore, the
generation of large quantities of waste is an issue that re-
quires attention due to high capital and operating costs
(Eccles, 1999).
After numerous studies, it is possible to confirm that the
most commonly used methods are high rates algal ponds
(HRAP) (Oswald, 1988; Mehrabadi et al., 2015). Furthermore, a
patented method known as Algal Turf Scrubber (ATS) was
developed to remediate AMD and polluted groundwater. It
uses suspended biomass of common green algae species such
as Chlorella, Scenedesmus, Cladophora or cyanobacteria such as
Spirulina, Oscillatoria, Anabaena or consortia of species (Craggs
et al., 1996; Adey et al., 1996). This structured approach was
patented as an Algal Turf Scrubber (ATS) in subsequent
U.S. Patents with the following details: 4,333,263 e 1982;
4,966,096 e 1990; 5,097,795 e 1992: and 5,851,398 e 1997. This
patented method has proved the effectiveness of this tech-
nology regarding the efficient removal of heavy metals to
acceptable levels in AMD or any other industrial wastewater,
and also an effective removal of organic compounds such as
chlorinated and aromatic organic compounds was recorded
(Craggs et al., 1996; Adey et al., 1996).
In a comparative study between a waste stabilization
ponds system (WSP) and HRAP for the treatment of urban
polluted water with lower ion concentrations of Zn, Cu and Pb
Toumi et al. (2000) reported that the HRAP had a higher
removal rate per unit volume per day with values up to 10
times more efficient in the case of Cu. High pH values were
recorded and they enhanced metal precipitation; the high pH
65
values are the consequence of effective algal photosynthesis.
A hybrid process that led to a patent was reported by Rose
et al. (1998), the patent emerged from a combination of
HRAP and SRB. In this process, the precipitation of heavy
metals is achieved by direct input of AMD into HRAP with high
pH values and the biomass having adsorbing properties. In
addition, the biomass of the HRAP is recovered and used as a
carbon source for SRB (Perales-Vela et al., 2006).
For effective removal of Mn and stability of a neutralizing
pH, non-agitated algal ponds have been suggested to reme-
diate challenges that constructed wetland technologies are
facing (Phillips et al., 1995). In this study it was shown that
consortia of algae and cyanobacteria could effectively reduce
prohibitive Mn concentrations to an environmentally safe
level. In this case Mn was removed by means of biomass
adsorption, high pH precipitation and immobilization
(Perales-Vela et al., 2006). All these previous studies point to a
promising future for algae bioremediation based treatment for
AMD and wastewaters.
Algae can produce oxygen at a lower costs and it is also
very effective in the removal of pollutants through phycor-
emediation (Wang et al., 2016). The existence of various spe-
cies creates opportunities that can be explored for treatment
of AMD. Spirulina sp. has been very effective regarding the
removal of heavy metals from AMD. Also, there is a variety of
acidophilic organisms growing easily in AMD but there is little
literature about their use for AMD treatment (Das et al., 2009;
Sanchez-Andrea, 2014). The ability of aquatic plants to remove
heavy metals efficiently from contaminated acid mine
drainage water can be traced back to 1973 (Dinardo et al.,
1991). Several algae strains have been successful in removal
of heavy metals from AMD or wastewaters, but most of the
investigations were completed on batch process in which
microalgal species are grown.
The function of algae as essential components of a wetland
with regard to remediation of AMD cannot be overlooked as
heavy metal contamination sinks through bioaccumulation.
This function plays a major role in the design of passive
bioreactor systems for the removal of sulphates in AMD
(Zagury et al., 2007). A passive bioreactor system operates on
the same principle as a large activated carbon filter with AMD
coming into the surface area, percolating through a specially
constructed barrier layer and exiting the system from below.
Algae and other aquatic plants are major components for the
initial stage of the bioreactor. Both Van Hille et al. (1999) and
Balaji et al. (2014) described the use of Spirulina sp. in the
remediation of AMD effluent; they discovered that rapid up-
take of various heavy metals through direct contact with the
AMD effluent stream reached saturation in 30 min. After this
time the algae began to exhibit toxicity effects. However, these
species of algae continuously generate alkaline chemical
materials that act to neutralize the acidity of the AMD through
the production of inorganic bicarbonate salts.
The effective process is chemically based and similar to an
acidebase neutralization reaction, rather than biologically
dominated where the uptake and sequestering of heavy
metals occurs directly in the algae. It is necessary to maintain
physical separation of the algal colony from the effluent
stream in order to prevent the algae from being affected by
heavy metals toxicity. Consequently, the alkalinity produced
by the Spirulina can continuously neutralize the acidic nature
of AMD effluent. The system is reported to have consistently
removed almost all heavy metals as recoverable hydroxide
compounds from the treated AMD.
66 south african journal of chemical engineering 24 (2017) 62e70
Many studies have been carried out to identify species of
both microalgae and macroalgae that can be applied to phy-
coremediation. A marine screen designed by Matsunaga et al.
(1999) was used to analyze a chlorella strain that was capable
of sustaining growth at 11.24 mg of Cd2/l with a removal ef-
ficiency of 65% when exposed to an amount of 5.62 mg of Cd2/
l. A trial using a batch process was initiated by Travieso et al.
(1999) using Chlorella and Scenedesmus strains at 20 mg Cr6/l. It
was found that the removal efficiencies were 48% and 31%
respectively for both strains.
Another strain known as Phaeodactylum tricornutum
(CE50 22.3 mg/l) with high Cd2 tolerance was characterized
with respect to Mt III production pattern by Torres et al.
(1997) and showed to have the potential to remove these
two metals (Torres et al., 1998). Scenedesmus was also used in
heavy metal removal studies and has demonstrated an
effective removal efficiency for U6 (Zhang et al., 1997), Cu2
and Cd2 as confirmed by Terry and Stone (2002) and Zn2
~ izares-Villanueva
(Aksu et al., 1998; Travieso et al., 1999; Can
et al., 2001).
In a study carried out by Freitas et al. (2011) on the bio-
sorption of heavy metals by algal species in AMD from coal
mining in Brazil, it was found that algal biomass are able to
accumulate heavy metals more specially Fe which constitutes
6.3% of the biomass. Their study showed the following ab-
sorption capabilities: Fe > Al > Ca > Mg > Zn > Mn > Cu in all
sampling sites. In addition their results also show that algal
species such as Microspora, Eunotia, Euglena, Mougeotia, and
Frustulia can survive in the AMD environment, with Microspora
being found to be the most dominant. These acidophilic spe-
cies were found to survive in water at pH values ranging from
2.9 to 4.1.
Park et al. (2013) completed a study using a hybrid system
comprising a pipe inserted microalgae reactor (PIMR) and an
active treatment unit for the removal of Fe from AMD. The
removal of Fe is very important because high levels of Fe can
be detrimental to algae growth and lipids accumulation,
thereby limiting the treatment ability of algae biomass when
dealing with AMD. In their study, it was found that Neph-
roselmis sp. is effective in removing heavy metals and has a
strong tendency to grow with pretreated AMD. Therefore, they
recommended that PIMR is more effective for both metals
removal and microalgae growth in AMD pretreated effluents.
Compared to HRAP and SRB, this system is more protective of
its algae biomass against environmental contaminations even
though the area of operation between algae biomass is
smaller. However, the treatment capacity and removal could
be less compared to the HRAP system.
Ben and Baghour (2013) reviewed at least fourteen algal
species and their ability for uptake of heavy metal contami-
nants from AMD and other pollution sources. In their study
individual species have showed to be hyper-accumulators or
hyper-absorbents with a high selectivity for different
metals. Also, Das et al. (2009) compiled some results achieved
by few studies regarding the use of some microalgae species
for AMD bioremediation. This table contains the removal ef-
ficiencies of heavy metals and sulphates which are the main
contaminants in AMD. It also includes the operating condi-
tions and the growth mode used to achieve the results. The
table indicates the effectiveness and possibility of algae
technology for AMD bioremediation. The main elements
which are heavy metals such as Cu, Zn, Fe, Pb, Ni, Mn and
sulphates can be removed by microalgae species under a
known pH range.
This specificity suggests the possibility of tailoring algal
phycoremediation processes to suit specific contaminant
conditions, as well as the possibilities for enhancing both
uptake limits and uptake specificity through genetic engi-
neering of algal species. The down side of active approaches
involving chemical, biological and microbiological processes
is the issue of clogging due to metals precipitation which
hinders the biological and microbiological activities, as
confirmed by Kalin et al. (2006). Table 1 presents some per-
formance data collected from studies that have used some
algae species regarding AMD bioremediation.
5. Discussion
The use of algae for the removal of heavy metals from AMD or
other contaminated wastewaters appears to be a common
component of passive water treatment systems, and has been
in use since the early 1990s. It has proved to be efficient for the
removal of both organic and inorganic contaminants, even
though there is still room for more studies to explore its full
potential with the development of innovative technologies.
This is particularly relevant to gene modification technology
which has the ability to identify specific genes responsible for
producing metal-chelating proteins and enhancing this
capability in algal strains.
Studies on algae and their biochemical mechanisms of
metal detoxification is growing and is widely accepted but
significant contributions are yet to be produced in light of the
success of current passive systems. Most studies have focused
on the growing field of biodiesel/biofuel production from algal
sources. This has overshadowed the potential value of algae in
phycoremediation which presents a great potential for AMD
remediation. This is supported by the use of the available
successful alkalinity producing systems (SAPS) methodology
and similar structures made up of natural environmental
materials and processes for efficient removal.
Some reported results appear to have exceeded perfor-
mance expectations with regard to water purification. At the
same time, however, maintenance problems such as clogging
of water flow by bio-organic materials and chemical precipi-
tation are being identified, as mentioned before. This requires
adjustment, monitoring and regular maintenance of the
existing systems in order to prevent or to overcome such
problems when they occur.
According to Das et al. (2009), more research should focus
on the use of Lepocinclis sp. and Klebsormidium sp. for AMD
treatment as they show promising features regarding their
growth and distribution. It was also recommended that algae
and fungi can be used as consortium in AMD either when used
as groups or individuals. They have an advantage of working
in symbiosis and synergy. Also, high concentration of heavy
metals, acidity, low availability of organic carbon and phos-
phates are detrimental to the growth of algae and fungi. This
can be challenging for an efficient treatment process because
AMD contains high levels of heavy metals with a very low pH
which is an indication of high acidity.
Compared to studies mentioned before, specially the one
completed by Ben and Baghour (2013), in which there was a
successful removal of heavy metals using algae based treat-
ment despite the high levels of heavy metals or acidity present
in AMD. The study completed by Das et al. (2009) focused on
specific species such as Spirulina sp., Lepocinclis sp. and Kleb-
sormidium sp. thereby limiting the scope of their study.
Therefore, for a conclusive study it is important to assess
 south african journal of chemical engineering 24 (2017) 62e70 67

Table 1 e Summary on use of some species in AMD bioremediation (adapted from Das et al. 2009).
Algal Species Remediation role
Blue-green algaee
cyanobacterial mat
Spirulina sp. Metal adsorption, re-
alkalization, nutrient for
SRB (dead biomass)
Spirulina sp. Alkalinity generation and
metal precipitation
Mixed algal population Soluble EPS as carbon
source for SRB
Eunotia exigua and Primary production
Pinnularia obscura
Spirulina sp. Dead biomass as nutrient
for SRB
Blue-green algae SO4 removal, metal
(predominantly precipitation by consortium
Oscillatoria spp.). e
microbial consortium
Chlorella ellipsoidea Bioremediation potential
Eunotia exigua and Enhance primary
Chlamydomonas sp. production thereby SRB
growth
Ulothrix Metal absorption
Achieved results Growth method Reference
Removal of 2.59 g of Mn/ Oxidation pond Phillips et al.
day/m2 (1995)
(i) Removal of Fe (up to HRAP (high rate algal pond) Rose et al.
100%), Zn (86e98%), Cu (38 (1998)
e76%), Pb (40e78%) at
retention time of 10 days.
(ii) Rise of pH from 3 to 8.5
for a biomass loading of
3 mg/mL chlorophyll a.
(i) pH rise from 1.8 to 8.18 Bench scale anaerobic Van Hille et al.
(ii) Reduction of sulphates digester, primary and (1999)
(SO4) 89%, Fe 99%, Pb 95%, secondary treatment.
Zn 93%, Cu 94%.
Up to 57% of sulphates (SO4) High rate algal pond (HRAP) Molwantwa et al.
and 52% COD removal by (2000)
mixed SRB
Chlorophyll a (Chl a) Mining Lake Koschorreck
content 52e72 mg/m2 et al. (2002)
150 mg SO4 removal/g algal Bench scale anaerobic up Boshoff et al.
biomass/day flow reactor (2004)
(i) pH increase from 2.93 to Bench scale test cell Sheoran and
6.78 Bhandari (2005)
(ii) Reduction of SO4 29%, Fe
95%, Pb 88%, Zn 86%, Cu
97%, Co 83%, Ni 62%, Mn
45%
In situ test using Mitmann et al.,
limnocorrals for EPA (2005)
Reduction of Fe from 14 mg/ Microcosm experiment Fyson et al. (2006)
L to 0.2 mg/L and SO4 from
344 mg/L to 124 mg/L
Absorption of Cu 3500 mg/L, AMD, Sar Cheshmeh copper Orandi et al.
As 500 mg/L mine (2007)

various algae species, their removal mechanisms and their
growth, pH, temperature, residence time and flow rate of the
AMD. A large variety of algal species exist naturally; however,
they can be classified as extremophiles, both requiring and
thriving in unusual or extreme environmental conditions.
Therefore, there is a need to examine in detail their adapt-
ability to be used in phycoremediation processes more espe-
cially for AMD.
However, it is important to state that in comparison with
conventional wastewater treatment plants, algae as sources
of oxygen have some weaknesses such as their unpredictable
response to a complex changing environment or wastewater.
Furthermore, critical environmental factors such as opacity of
the medium, solar radiation, rise of nutrients, temperature,
season and ecological succession are not easily understood
and they need to be monitored meticulously for the success of
the process.
6. Shortcomings of phycoremediating using
aquatic plants
Phycoremediation is a better emerging environmentally
friendly and cost-effective method for the treatment of
wastewater and polluted environments (Sharma et al., 2014;
Sood et al., 2012; Emmanuel et al., 2014; Newete and Byrne,
2016) when compared to conventional methods. Its ability to
grow fast and to produces a biomass is an advantage while its
poor tolerance to high metal concentrations and its seasonality
is a disadvantage to this treatment technology (Rai, 2008;
Mannino et al., 2008).
Due to the high concentration of heavy metals and their
toxicity, phycoremediation is used as either a secondary or
tertiary treatments method for industrial or mine wastewater
treatment (Sharma et al., 2014). So for effective phycor-
emediation treatment, there should be regular harvest and
safe disposal of the biomass (Rai, 2008) to avoid the release of
the absorbed or adsorbed metals back into their source when
the plant dies and decompose.
The safe disposal of phycoremediation aquatic plants is an
emergent area of research that needs to be addressed
(Newete and Byrne, 2016). The use of hyper accumulating
aquatic plants such as algae for treatment technology of
arsenic has an environmental and promising economic
prospects but the accumulation and removal of the metalloid
is not enough for the successful implementation of this
technology if it is not effectively managed and disposed
(Rahman and Hasegawa, 2011)
7. Unsolved issues and novel approach
They are currently many unsolved issues that algae based acid
mine drainage treatment may face currently. Among them is
the identification of a very efficient species that can have a
very high active uptake capacity for heavy metals including
other soluble contaminants. The species should be able to
thrive in a very acidic media. Algae species are temperature
68 south african journal of chemical engineering 24 (2017) 62e70
sensitive and depend on natural light energy from the sun for
growth. They are more acclimated in warm or tropical envi-
ronment however, in cold environment with low natural solar
energy, their operability and capacity to remediate acid mine
drainage can be a great challenge.
Mostly, the flow rate of acid mine water is very high in
various mines, therefore, there is a possibility to generate high
quantities of contaminants from the effluents. Consequently,
high amount of biomass will be needed for the removal of
contaminants. This will require the cultivation of algae in
large area or development of very efficient cultivation system
to be able to solve a continuous flow of AMD.
The cost of separation by clarification or harvesting algae is
still not satisfactorily up to date. Many processes are used for
harvesting the biomass, some are energy intensive and other
are time consuming. The new approach will be to develop a
hybrid system that combines the conventional treatment
process with algae treatment incorporated as secondary
treatment. In this approach algae will bring oxygen and
remove heavy metals during the treatment. The tertiary
treatment will carry on as per normal in a conventional
treatment using precipitation and disinfection or any other
chemical reactions to remove the remaining contaminants.
This option can be energy saving because oxygen can be
supplied by microalgae, however, the costs for algae har-
vesting can be another challenge, therefore, optimization of
the harvesting process needs to be undertaken.
Harvesting remains the best way to accomplish an effec-
tive separation between the clean water and biomass. Sedi-
mentation of algae is not easy because microalgae size are
very small, they are expressed in micron, the settling is very
challenging unless using harvesting techniques such as
filtration, coagulation and flocculation or centrifugation.
8. Challenges and future prospective
Excluding environmental factors, there are various issues
related to biological and operational aspects which include
contamination, auto-inhibition and grazing taking place during
the growth of microalgae for wastewater or AMD treatment.
These issues may affect seriously the treatment performance
of AMD or wastewater when using algae based treatment.
Therefore, daily monitoring of algae growth, protection of the
growing medium, sterilization and ultra-filtration of the cul-
ture medium which can be an open pond are the most
important actions that are needed to be undertaken.
The main weakness of algal open pond is that it requires
larger space compared to other forms of sewage treatment. In
this case, it is important for design purposes to consider local
land prices and soil suitability in selecting the least cost
method if dealing with algae based treatment of AMD or
wastewater. In many cases, ponds are considered as the best
choice for the treatment, as the land is habitually accessible at
affordable costs.
Contamination by bacteria can be easily treated with the
use of appropriate chemicals. Studies need to be undertaken to
address issues related to contamination and auto-inhibition.
Also, the choice of the specie is very important because not
all species can treat AMD efficiently. Many studies have
demonstrated the capacity of algae to remove heavy metals
and many other compounds, however they are still at the
laboratory level, there is need for scaling up the technologies to
help solve the issue related to management and treatment of
AMD which has not been solved efficiently till date.
9. Conclusions and recommendations
Literature and studies from various sources indicate that the
use of algae in phycoremediation is effective and has potential
for future applications as indicated in this current review. It
was also noted that the composition of AMD varies from site
to site due to different pH, temperatures and oxygen content
values on each site. These variations imply that different
species of acidophilic microorganisms with different physio-
logical characteristics will be found on each site where AMD is
generated. This knowledge will shed more light on the
microbiology of AMD and can lead to a better understanding
of the biogeochemistry of AMD environments.
Particular contributions such as the creation of sulphates-
reducing anaerobic regions in passive water systems greatly
increase the effectiveness of those systems in removing heavy
metals from AMD and other contaminated wastewaters.
These systems have a particular advantage of generating
higher alkalinity in the effluent to facilitate precipitation of
the entrained metals as poorly soluble but economically
recoverable oxide and hydroxide salts.
Other new and effective technologies, such as reactive
permeable barrier systems are also identified as potentially
effective for future application dealing with removal of heavy
metals in AMD. The current success is overshadowing these
systems, also the research and development of highly effec-
tive barrier materials (natural and synthetic) has been limited.
From all studies reviewed, the universal nature and the
seriousness of AMD and other associated heavy metals
contamination were clearly pointed out. It is also acceptable
that the current passive systems, while achieving commend-
able results, are not the ultimate solution to the problem.
Therefore, more research on effective means of remediation is
recommended to help solve the environmental challenges
brought by AMD.
It is a fact to state that each individual site contaminated
by AMD has its own unique characteristics and requirements
for remediation. More research on different aspects of algae
based treatment should be undertaken. This should range
from identifying and developing simple, effective physical
barrier materials to advanced genetic research suitable for
AMD.
Acknowledgments
The authors wish to extent their gratitude to the research
directorate of Mangosuthu University of technology for the
support.
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