BIOFILM REACTORS IN MINING AND METALLURGICAL EFFLUENT

TREATMENT: BIOSORPTION, BIOPRECIPITATION, BIOREDUCTION

PROCESSES

M. Tsezos, A. Hatzikioseyian, E. Remoudaki

National Technical University of Athens,
School of Mining and Metallurgical Engineering,
Laboratory of Environmental Science and Engineering,
Heroon Polytechniou 9, Zografou 15780, Athens, Greece.
Tel +30 210 772 2271 Fax +30 210 772 2173,
e-mail: tsezos@metal.ntua.gr

ABSTRACT

During the last decade, considerable volume of work has been carried out by
Institutes, Universities and private corporations on the development of novel
biotechnological processes for the advanced treatment of metallurgical and mining liquid
effluents. Reactors that can support the development and maintenance of biofilms, such
as, static beds, fluidized beds, moving bed sand filters and rotating biological contactors
(RBC), provide a variety of liquid effluent advanced treatment options for the above
categories of effluents. Biofilm reactors, both in pilot and industrial scale, have been
tested and the available results on pollutant sequestering are very promising. The
successful development of biofilm reactors has been based both on the research advances
on the characteristics of microbial biomass species and on the elucidation of the likely
mechanisms of interaction between the microbial biomass and the targeted pollutants
which include toxic metals, radionuclides and metalloid oxyanions. Microbial biomass-
metals interactions occur through a variety of mechanisms. These mechanisms can be
classified in two principal categories: the metabolically active (biomineralization,
biotransformation, bioprecipiation) and metabollically passive (biosorptive) processes.
Metabolic interactions between microbial biomass and metals or oxyanions in liquid
effluent solutions may lead to metals precipitation (bioprecipitation), metals reduction
(bioreduction) etc. Passive microbial biomass-metal/metalloids interactions include
extracellular, intracellular sequestering, chemical bonding, complexation and ion
exchange mechanisms. This variety of metals sequestering mechanisms by microbial
biomass, together with the low amounts of energy and materials required for their scale
up and application, make biotechnological processes promising for efficient metal

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removal and recycling at waste water polishing stage. The present paper summarizes the
main process characteristics and some of the results available both in pilot and in
industrial scale applications, including a list of references for further study on the subject.

INTRODUCTION

Biological methods for removal or recovery of heavy metals, radionuclides and

oxyanions, have been developed during the last three decades as polishing stage in a
wastewater treatment scheme. Main advantages of these methods are:
• Environmentally friendly concerning energy and material consumption
• Possibility of metal recycling and recovery
• Reduced amount of sludge production for disposal
The typical problem of the effluents generated in the mining and processing of metal ores
is the large volume of wastewater and process water very often characterised by acidity
and high content of sulfates and metals. The most commonly applied technology to
remove the contained metals is precipitation with lime or other agents. This leads to the
precipitation of metals as hydroxides, which are then typically landfilled. The operating
and disposal costs of this approach are high, while the concentrations of residual metals
and sulfates are in the orders of 1500 mg/l for sulfate and 0.5-5 ppm for the metals. In
addition any valuable metals present in the waste stream are lost in the solid phase
sludge.
Biological methods implemented using various types of biological reactors aim at
sequestering of metals, radionuclides and metalloids from mining and metallurgical
effluents. Novel biotechnological methods for industrial wastewater treatment are based
on the development and maintenance of specific active biofilms on an appropriate solid
support medium.
Interactions between the targeted pollutants in the effluent to be treated and the
biofilm can occur in number of mechanistic pathways such as:
(a) Microbial accumulation. Passive or active accumulation of metals by microorganisms
can serve as means of metal immobilization. Microorganisms possess an abundance of
functional groups on their cell surface and extracellular polymers that can passively
sequester metal ions. This process is known as biosorption and has been extensively
studied, (1-8).
(b) Microbially mediated precipitation. This process involves the microbially induced
precipitation of metals and radionuclides as minerals of sulfide (9-11), hydroxide (12,
13), phosphate (9) and carbonate (14). Microbially mediated precipitation is important in
the geological cycle of the elements in nature, and has potential as well as documented
cases of application in industrial wastewater treatment technology and bioremediation.
These processes are commonly referred to as biomineralization or bioprecipitation
processes. For the case of sulfide bioprecipitation, the biological reduction of sulfate to
sulfides by Sulfate Reducing Bacteria (SRB) is considered one of the most important
environmental biotechnological applications. Sulfate reducing bacteria belong to the
genera Desulfobacter, Desulfococcus, Desulfosarcina, Desulfobacterium, Desulfonema,

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Desulfovibrio, Desulfotomaculum, Desulfomonas, Thermodesulfobacterium and
Desulfobilbus. (15).
Another bioprecipiation example, is the enzymatically produced HPO42- by Citrobacter
sp. which has been shown to precipitate metals as phosphates, (16-18).
Some other microbial products, such as microbial exudates induced by the presence or
absence of specific metal ions, e.g., iron binding siderophores and toxic-metal binding
proteins and extracellular polymeric secretions (EPS), are also responsible for metal
complexation and immobilization, (19, 20).
(c) Microbial transformation. Metals or metalloids can be transformed via reduction /
oxidation or alkylation reactions by microorganisms. For example, many aerobic and
anaerobic bacteria reduce Cr(VI) to the less toxic and less soluble Cr(III).
The bioreactor conditions (i.e. aerobic, anoxic, anaerobic etc.), the type of
biomass (aerobic, denitrifying, SRB, active or inactive biomass etc) and the effluent
composition (i.e. presence of nitrate, sulfate etc.) are key factors determining the
dominant mechanism in altering the targeted element mobility or speciation, whereas,
other secondary mechanisms, among those mentioned above, may also contribute to the
observed pollutant sequestering from the effluent under treatment.
A key advantage of fixed film processes is that high densities of microorganisms
per unit volume of reactor can be supported through their natural attachment as biofilms.
This high density of biomass accumulation allows excellent treatment performance in
relatively small reactor volumes, which is economically advantageous. Another
advantage is the fact that the need for sedimentation is limited. Normally there is no need
for sludge recirculation and sometimes there is no need for recirculation of the effluent
(21, 22). The excellent biomass retention and relatively short hydraulic detention times of
biofilm processes make them attractive when bacterial growth rates are slow or when the
compounds are slowly degraded or inhibitory, as is the case of the metal bearing
industrial effluents. The immobilized biomass offers the advantage of surviving from
shock loads of toxic wastes due to the relatively short retention time of the wastewater in
the reactor, which means that only organisms on the surface of the biofilm may be
affected (23).
Because of the above advantages, during the late 1980s and 1990s considerable
research and development effort was directed towards novel biofilm reactor systems and
applications. Systems such as small-granule fixed-beds, fluidized beds, rotating
biological contactors, trickling filters, and hybrid suspended biofilm processes were
developed.
The present paper briefly reviews the most recent pilot and industrial applications
of immobilized biomass reactors used in mining and metallurgical wastewater polishing
treatment, presenting necessary background information and key concepts of the
technology.

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 RESULTS FROM PILOT AND INDUSTRIAL APPLICATIONS

Biosorption

Pilot installations and few commercial scale units were constructed in the USA
and Canada during 1980s and 1990s, (see Table I). These pilot plants confirmed the
applicability of biosorption as the basis for metals sequestering/recovery process
especially in the case of Uranium where it was tested in order to be combined with in situ
bioleaching, giving rise to an integrated biotechnology based Uranium production
scheme, (29). These pilot plants helped the researchers to realize limitations associated
with the use of inactive microbial biomass in the industrial application of biosorption,
mainly due to the cost of formulating the biomass into biosorbent material. Furthermore,
the negative effect of solution matrix co-ions, on the targeted metals uptake by the
immobilized microbial biomass and the reduced resilience of the biological material,
made recycling and reuse of the biosorbent even more difficult, (30).

Table I. – Biological basis sorbent particles (Biosorbents) developed for metal bearing
wastewater treatment applications, (32).

Immobilization
Name Microorganism Particle size References
matrix
Cyanobacteria
Polyethylene or
Bio-fix (Spirulina)
Polypropylane or
U.S. Bureau of Mines, Yeast 0.5-2.5 mm
Polysulfone in 24
(Golden, Colorado) Algae
dimethylformide
Plants (Lemna sp.,
Sphagnum sp.)

AMT-Bioclaim Bacillus subtilis 25

AlgaSORBTM
Silica or
Bio-recovery system Inc., Chlorella vulgaris 26, 27
polyacrylamide gels
(Las Cruces, New Mexico)
Sargassum natans
Ascophyllum
nodosum
B.V.Sorbex Inc.,
Halimeda opuntia 6
(Montreal, Canada)
Palmyra pamata
Chondrus crispus
Chlorella vulgaris
Rhizopus arrhizus
Polymer
Tsezos Activated Sludge 0.5-1.0 mm 28
coating
P. chrysogenum

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Biosorption however is a process with some unique characteristics. It can effectively
sequester dissolved metals out of very dilute complex solutions with high efficiency.
This makes biosorption an ideal candidate for the treatment of high volume low
concentration complex wastewaters. It has recently been shown that in cases of
metabolically active microbial cells, in biological reactors, biosorption contributes as a
parallel mechanism together with other metabolically mediated mechanisms such as
bioprecipitation and bioreduction, (31).

Bioreduction

The microbial transformation of metals has also attracted interest, as the

microbial metal reduction covers a wide range of metals including: Fe(III), Mn(IV),
Cr(VI), Hg(II), Co(III), Pd(II), Au(III), Ag(I) Mo(VI), V(V), some metalloids such as
As(V) and Se(VI) and radionuclides U(VI), Np(V) and Tc(VII), (33). The reduction of
these elements serves for (a) conserving energy for growth e.g. Fe(II) or Mn(IV), (b)
reducing cell’s environment potential toxicity e.g. Cr(VI), Hg(II), As(VI) or (c) proceed
enzymatically under non-energy conserving metal-reductase activity e.g. U(VI), Cr(VI),
Tc(VII), Mo(VI), Pd(II), (34). The microbially mediated reduction of these elements
commonly results in species of lower toxicity and mobility. Both characteristics are key
targets for successful soil or water bioremediation technology.
Large-scale applications of metal bioprecipiation - bioreduction are mainly
applied in contaminated soil and groundwater bioremediation projects, (35, 36).

Figure 1. Laboratory scale Rotating Biological Reactor (RBC) for treatment of Cr(VI)
containing effluents through bioreduction to Cr(III) and bioprecipitation,
(Courtesy, NTUA, Laboratory of Environmental Science and Engineering).

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 A European Project on the biological reduction of oxyanions such as chromate
(CrO42-), arsenate and selenate examined possible microbial technologies for the removal
of Cr, As and Se from wastewater, (37). The results of this project revealed that
biological reduction of the oxyanions is successful mainly in the anaerobic zones of
treatment systems such as packed bed sand filters and rotating biological contactors. As
an example of biologically reduced Cr(VI) treatment application, a small scale unit with
immobilised biomass in a Rotating Biological Contactor (RBC) unit has been tested and
reported, (31, 37). The reactor, with a total volume of 100 L, had 46 removable
polyethylene disks, each with characteristic hexagonal pyramidal dimple structures in
order to increase the support surface for microbial immobilization, (Figure 1). The unit
was inoculated with microbial strains capable of tolerating increased concentrations of
Cr(VI) and Cr(III) and having the ability to reduce chromate ions. The system operated
for more than 2 months with complete reduction of Cr(VI) to Cr(III) for influent
concentration ranging from 4 –10 ppm Cr(VI), (31, 37). The resulting Cr(III) could be
precipitated under specific conditions, by adjusting the pH around circumneutral values
and by maintaining the excess of the nutrients or complexing agents in low levels.

Bioprecipitation

Microbial precipitation of metals and radionuclides as minerals of sulfide,

hydroxide, phosphate and carbonate, has been documented in full-scale applications in
wastewater treatment technology and bioremediation. The term applies to biological
precipitation mechanisms also known as biomineralization or bioprecipitation. Two
novel efficient technologies based on bioprecipitation are briefly presented below.

Moving bed sand filters

A novel type immobilized biomass biological reactor, in a moving bed sand filter
configuration, has been evaluated in pilot scale for the removal of heavy metals in
mining, metallurgical and plating industrial effluents in the frame of a European
Research Project, (38). The operation of the filter is based on the fact that sand grains act
as support material for biomass immobilisation. The microbial biofilm formed interacts
with the soluble metal species of the wastewater treated. Key characteristic of three
reactor is that the sand is continuously circulating, washed, and the produced excess
biomass is continuously removed, (39). Figure 2, schematically shows the operation
principle of these reactors. The operation of the filter can be described as follows, (39):
WATER: the effluent to be treated enters the filter by the pipeline [1]. The water enters
the filter bed [4] through the supply pipe [2] and the distributors [3]. The water is treated
as it flows through the filter in upward direction. The treated water is discharged in the
upper part of the filter [5].
SAND: the filter bed is continuously moving downwards as the water flows up. The sand
with the immobilised biomass is abstracted from the sand bed [6] and washed [10] after
which the sand is released back on the top of the sand bed, [7].

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 Figure 2. Schematic diagram of a moving bed sand filter, (adopted by 39).

AIR: The sand circulation is based on the airlift principle, forcing a mixture of biomass,
sand and water upward trough a central pipe line [8]. The intensive scouring movement
separates the biomass from the sand particles. At the top of the pipe line the air is
released and the wash water is discharged among with the metal loaded biomass [9]. The
washed sand is settled in the washer [10].
WASHER: The washer [10] is position around the airlift pipeline. The sand particles fall
through the washer where they are finally washed by a small amount of filtrate, flowing
through the washer countercurrently. The filtrate flow is generated by a difference in
discharge level between the filtrate [11] and the washwater [9].
The operation of the pilot moving sand bed filters in mining, metallurgical and
plating industrial applications have shown very encouraging results concerning metals
removal at sub-ppm levels, (38, 40). In pilot applications, the biomass growth was
supported by the addition of selected carbon sources. Acetate is a carbon source that
when metabolized by microbial biomass generates alkalinity, which alters the chemical
microenvironment of the biofilm, by increasing the pH and carbonate/bicarbonate
concentration of the growth medium. Targeted metals present in soluble forms in the
wastewater, are retained in the biological sludge generated during the operation of the
sand filters. The experimental results show that metals precipitation takes place by the

285
cells microenvironment and is favored by the metabolically generated alkalinity. This
phenomenon can be described as metal bioprecipitation.
During the sand filter operation, the development of an anoxic zone following the
aerobic zone at the feed level, offers the possibility of metal precipitation in both zones.
Different insoluble metal compounds are formed in each zone. Acetate microbial
metabolism in the aerobic zone leads to the formation of metal carbonates and
hydroxides. In the anoxic zone and in the presence of sulfates, metal sulfides are formed
which are the most insoluble among the simple metal species, which could be formed.
This is a superior advantage of processes operating under anoxic sulfate reducing
conditions for metal ion wastewater treatment.
Precipitated metals in the produced biological sludge are composed from more
than one insoluble species. These species depend mainly on: (I) the metal composition of
the feed wastewater, (II) the biological conditions prevailing during the sand filter
operation (development of anoxic zone) and (III) the presence of alternative electron
acceptors such as sulfates in the influent, (40).

THIOPAQ® technology for sulfate reduction and metal removal by sulfate reducing
bacteria (SRB)

The core of the technology is the biological reduction of sulfur compounds, which
are reduced to sulfide (H2S), and the subsequent precipitation of metals. This is described
below by the following equations:

SO42- + 8 H+ + 8e → S2- + 4H2O (1.1)

Me 2+ 2-
+S → MeS (1.2)

The first reaction is carried out by sulfate reducing bacteria in a biological

reactor. Besides sulfate, practically all other oxidized sulfur compounds (SO2, sulfite,
thiosulfate, (poly)sulfide, elemental sulfur etc.) can be biologically reduced to sulfide.
For the reduction of sulfur, hydrogen or other organic compounds (i.e. ethanol, methanol,
lactate, waste products, etc), can be used as electron donors.
The solubility of metal sulfides (ppb levels) is lower than the solubility of the
corresponding metal hydroxides, thus lower metal discharge standards can be met as
compared to lime treatment. Moreover, lime treatment cannot reduce the sulfate
concentration below 1500 mg/l, whereas biological treatment results in sulfate
concentrations of less than 300 mg/l. In addition, the metal sulfide precipitate is less
voluminous resulting in lower disposal costs. The precipitated metal sulfide / sulfur
sludge can be recirculated to the front-end of a sulfide smelting or refining operation.
Based upon the principle that the solubility of metal sulfides is determined by the
pH value, metals can be selectively recovered. For instance it is possible to precipitate
separately copper, arsenic and zinc. The second reaction is a physical precipitation
reaction, where metal sulfides formed are crystallized into larger particles, which are
removed by sedimentation.

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 When more sulfide is produced than the stoichiometric requirement for
precipitation of the metals, the excess sulfide can be converted biologically into
elemental sulfur with alkalinity production according to the following reaction.

HS - +1/2 O2 → S 0 + OH - (1.3)

The sulfur formed can be reused for sulfuric acid production or marketed as
fertilizer.
Applications of this technology can be implemented in a variety of industries for
any aqueous stream containing metals and/or sulfur compounds. The most common
applications are:
• Selective metal recovery for weak acid streams, process water, acid mine
drainage, smelter effluents, ground water, tailings bleed or leach streams etc.,
• Sulfate and thiosalts removal by conversion to elemental sulfur,
• Production of gaseous H2S for metal recovery in metallurgical and mining
operations, (the safe and cost effective alternative to liquid H2S and NaHS),
• SO2 removal (producing sulfur or H2S),
• Simultaneous removal of metals sulfates nitrates and organic compounds.
The latest sulfate reducing technology is implemented at Pasminco Budel Zinc in
the Netherlands, (41). The core of the process is a 500m3 sulfate-reducing gas-lift
bioreactor: The bioreactor is run with hydrogen gas as electron donor. In this reactor zinc
sulfate is precipitated as zinc sulfide according to the following reaction:

ZnSO4 + 4H2 → ZnS + 4H2O (1.4)

The metal sulfide and biomass are retained in the reactor by a settler located on
the top. The metal sulfides are allowed to build up from 10 to 20% in the bioreactor. The
excess is withdrawn, dewatered and returned to the zinc smelter. The effluent contains
very low concentration of heavy metals, sulfate and sulfides and is directed to the
existing ground water treatment plant for final polishing, (41).

CONCLUSIONS

Biofilm reactor configurations recently developed and tested both in

demonstration and full-scale applications offer significant advantages and have shown
very promising results for the treatment of mining and metallurgical effluents.
Bioprecipitation, bioreduction and biosorption may act simultaneously in such reactor
configurations.
Among the known biological reactor configurations, two novel technologies: (a)
moving bed sand filters and (b) THIOPAQ technology apply to a wide range of mining
and metallurgical effluents treatment, presenting outstanding advantages over traditional
biofilm packed bed columns and the classic physico-chemical treatment processes
satisfying the requested effluent standards.

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 In the moving bed sand filter, a variety of bioprecipitation products may be
formed depending on the characteristics of the sand bed zone (aerobic, anoxic,
anaerobic). Bioreduction of metals may also take place. In addition passive and/or active
biosorption contributes to the targeted elements sequestering. These advantages together
with the continuous operation self-cleaning mode of the filter and the high volumetric
flow-rate capacity make such biological reactors an attractive technology for most
mining and metallurgical liquid effluents treatment.

ACKNOWLEDGEMENTS

Part of the results appearing in this paper have been produced in the frame of the
following European Union projects:

BRITE EURAM III ΒΕ95-1610/BRPR-CT96-0172, “Removal and Recovery of Heavy

Metals from Waste Water by Sand Filters Inoculated with Metal Biosorbing or
Bioprecipitating Bacteria”, (1996-1999).

BRITE EURAM 96 3584 “Development and assessment of biotechnological methods for

the removal of the anions of arsenic and chromium from wastewater”, (1997-1999).

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