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HAZMAT-7481; No. of Pages 7 ARTICLE IN PRESS

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Journal of Hazardous Materials xxx (2007) xxx–xxx

Integrated bacterial process for the treatment

of a spent nickel catalyst
V. Bosio, M. Viera ∗ , E. Donati
Centro de Investigación y Desarrollo de Fermentaciones Industriales (CINDEFI), Departamento
de Quı́mica, Universidad Nacional de La Plata – Consejo Nacional de Investigaciones Cientı́ficas
y Tecnológicas (CONICET), 50 y 115, 1900 La Plata, Argentina
Received 9 April 2007; received in revised form 20 September 2007; accepted 29 October 2007

Abstract
Integrated biological processes involving the dissolution and subsequent precipitation have been used for the treatment of the spent material from
the hydrogenation of vegetable oil containing a high-level of nickel. Our results show that nickel was successfully leached using Acidithiobacillus
thiooxidans. The percentages of nickel leached using A. thiooxidans were higher than those obtained with dilute sulphuric acid solutions. Due to the
physical characteristics of the residue, the best results were obtained when the leaching process was carried out using sulphuric acid biogenerated by
an A. thiooxidans biofilm. The recovery of nickel from the leachates was performed at room temperature by precipitating with sulphide generated
by Desulfovibrio cells. Indirect precipitation using sulphide generated in Desulfovibrio sp. cultures allowed the recovery of nickel as the very
insoluble nickel sulphide.
© 2007 Elsevier B.V. All rights reserved.

Keywords: Nickel; Recovery; Spent catalyst; Desulfovibrio; Acidithiobacillus thiooxidans

1. Introduction Suitable catalysts for hydrogenation processes can be eco-

nomically produced using metallic nickel and other nickel
Several chemical industries use large quantities of solid cat- compounds. According to the Environmental, Health, and Safety
alysts containing valuable heavy metals such as Pt, Pd, V, Mo, (EHS) Guidelines for Vegetable Oil Processing published by
and Ni. Due to the gradual build up of impurities which even- the International Finance Corporation [2], nickel catalysts from
tually plugs the pores and deactivates the catalysts, they often hydrogenation should be either: (i) recycled and recovered for
require replacement after 2 or 3 years of operation [1]. Spent cat- reuse as a nickel catalyst, nickel metal, nickel salt, etc. or (ii)
alysts are managed through: (i) chemical recovery and recycling stored and disposed of according to the waste management guid-
of valuable metals for different applications, (ii) regeneration ance presented in the General EHS Guidelines. In most cases
for reuse and (iii) landfilling. The regeneration of spent cata- the spent catalysts are mixed with filtering aids (diatomaceous
lysts is only possible in a limited number of catalytic systems earth) and these mixtures should be treated in the same way
and can only be carried out a few times. Recycling of spent as proposed for nickel catalysts. Approximately 6–9 kg of this
catalysts lowers the catalyst cost as well as the environmental mixture is generated per tonne of oil produced. In our country,
pollution caused by catalyst waste. Landfilling for the disposal the spent nickel catalyst used in the hydrogenation of vegetable
of spent catalysts is becoming increasingly difficult due to both oil is mixed with diatoms but no further treatment is performed.
the decreasing availability of landfill space and the concern for Thus, this solid residue accumulates in heaps, representing a
pollution arising from possible leaching of heavy metals. potential hazard to human health and to the environment, and
also a waste of a valuable non-renewable resource.
Traditionally, metal recovery from spent catalysts has been
carried out by leaching with inorganic acids or smelting [3].
∗ Corresponding author. Tel.: +54 221 4833794; fax: +54 221 4833794. These processes entail the use of acids in large-scale operations,
E-mail address: [email protected] (M. Viera). which generate large volumes of potentially hazardous waste

0304-3894/$ – see front matter © 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.jhazmat.2007.10.095

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and gaseous emissions. Bioleaching processes are based on the was carried out using the sulphide biogenerated by Desulfovibrio
ability of microorganisms to transform solid compounds into sp.
soluble and extractable elements that can subsequently be recov-
ered. Bioleaching processes have been successfully applied on 2. Materials and methods
an industrial scale for the recovery of copper metal from natu-
ral ores and for the biooxidation pretreatment of refractory gold 2.1. Spent catalyst
ores [4]. This approach can be considered as a ‘clean technology’
and this is associated with lower cost and energy requirements The spent catalyst used throughout this work had been
in comparison to non-biological processes [5]. Bacteria from employed in the hydrogenation of vegetable oil. The waste
the genus Acidithiobacillus are the most important microorgan- material was an amorphous black mass consisting of the spent
isms applied to metal solubilisation, particularly because they catalyst, organic material adsorbed onto its surface (vegetable
are autotrophic and tolerate high concentrations of heavy met- oil, products from incomplete hydrogenation, polyhydric alco-
als. Cells derive the energy required for their metabolism from hols, and fatty acids) mixed with a filtering aid (diatomaceous
the catalysis of the aerobic oxidation of reduced sulphur com- earth).
pounds including sulphides, elemental sulphur and thiosulphate. The organic material adsorbed on the catalyst did not allow
Furthermore, Acidithiobacillus ferrooxidans can use Fe(II) as an adequate contact between the waste material and the solution.
energy source [6]. The usefulness of these microorganisms in As a result, one part of the spent catalyst was treated with two
metal solubilisation – not only from ores and concentrates but parts of xylene at 60 ◦ C during 1 h prior to use. The residue
also from solid wastes – is mainly related to their ability to gen- was washed with ethanol and dried at room temperature. This
erate acidic media (sulphuric acid is the last product during the procedure was repeated at least twice. The nickel content was
oxidation of sulphur compounds) and oxidising media [Fe(III) determined as follows. Two samples (0.1 g each) of the spent
is produced during the growth of A. ferrooxidans on Fe(II)]. catalyst were treated with xylene and then digested with HNO3
After metals are released from the solid wastes, they should and diluted to 50 ml with distilled water. The resulting samples
be recovered from the liquid phase. Precipitation with lime is were each filtered through a 0.45 ␮m membrane and the nickel
the conventional technique applied for the immobilisation of concentrations in the final solutions were determined. The nickel
metals. However, this treatment has some significant limitations concentration in the residue was 41 ± 2 mg of nickel per gram.
in terms of applications and effectiveness as it usually results
in production of mixture and unstable metal hydroxides [7].
2.2. Microorganisms
Microbial processes are especially suitable techniques for the
recovery of elements present in relatively low concentrations in
The strain of A. thiooxidans (DSM 11478) used through-
solution where conventional techniques are usually less effective
out this work was routinely maintained in iron-free 9K culture
[5,8]. Sulphate reducing bacteria (SRB) are heterotrophic bacte-
medium [17] with 1% (w/v) powdered elemental sulphur, as an
ria that grow under anaerobic conditions and reduce sulphate to
energy source.
sulphide. Most heavy metal sulphides have a very low-solubility
Sulphate reducing bacterium Desulfovibrio sp. (ATCC
product so that even a moderate output of sulphide can remove
49975) was used in the precipitation experiments. The strain
metals to levels permitted in the environment [9]. Several labora-
was routinely maintained in Postgate’s B medium [18] at 4 ◦ C
tory experiments have been carried out to study the possibility of
in 10 ml bottles sealed with a rubber stopper and aluminium cap.
using SRB to precipitate heavy metals from wastewater both in
situ and ex situ [10–12]. There are a few examples of processes
that utilise SRB in the treatment of contaminated water on an 2.3. Leaching experiments
industrial scale. The best known is the groundwater treatment
process installed at the Budelco Zinc Refinery (The Netherlands) 2.3.1. Chemical leaching experiments
since 1992 [13]. Chemical leaching tests were carried out with solutions of
Integrated processes that involve dissolution and subsequent sulphuric acid at different pH values (1.0, 2.0, 3.0 and 5.0). Solu-
precipitation can be used for metal recovery. These processes tions were prepared by adding concentrated sulphuric acid to
are based on the ability of Acidithiobacillus to leach metals distilled water. The leaching tests were conducted using 100 ml
from soils or solid wastes. The metals ions, which are released of the acidic solution in a 250 ml Erlenmeyer flask with 1% (w/v)
as soluble ions in a highly acidic solution, are later removed pulp density of spent catalyst at 30 ◦ C and 150 rpm. All exper-
by precipitation with sulphide produced by the activity of SRB iments were conducted in duplicate. Samples were collected,
[14–16]. filtered and analysed for soluble nickel and pH. Samples were
In this paper we present the results of an integrated process taken the following days: 1st, 2nd, 8th, 15th and 22nd.
aimed at the treatment of the spent nickel catalyst generated in
the hydrogenation of vegetable oil process. Nickel was leached 2.3.2. Bioleaching experiments
from the solid residue using A. thiooxidans cultures. The effects Bioleaching was performed in 250 ml Erlenmeyer flasks with
of several factors (pulp density, type of reactor, pH) were anal- 100 ml of iron-free 9K medium (pH 5.0), the spent catalyst
ysed. Abiotic leaching experiments with sulphuric acid were at various pulp densities (0.2, 1 and 2%, w/v) and 1% pow-
also performed. The recovery of nickel from the acidic solution dered elemental sulphur. Flasks were inoculated with 10% (v/v)

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cells, harvested by filtering the grown culture with blue rib- Table 1
bon paper followed by centrifugation (20 min, 2000 × g) and Conditions used in the experiments of bioleaching with sulphuric acid generated
by an A. thiooxidans biofilm
then suspended in fresh medium. The centrifugation–suspension
procedure was repeated until the pH was ca 5 (washed cells). Volume of leaching pH Recirculation rate Spent catalyst (g)
In order to overcome the inhibition in the bacterial growth due solution (ml) (ml/min)
to the alkalinity of the spent catalyst, two other sets of experi- #1 143 0.66 5 10
ments were designed. Firstly, bacterial cells from a grown culture #2 200 0.52 5 15
(unwashed cells) were directly inoculated to the flask (prepared #3 150 0.62 10 15
#4 160 0.8 13 15
as indicated above). In this way, the acidity of the inoculum
avoided the rapid increase in pH and the consequent inhibition The volume and pH of leaching solution (generated in C1 by the A. thiooxidans
of bacterial. In the second set of experiments, flasks were pre- biofilm) pumped to the C2 at a constant rate of 113 ml/min; the recirculation
rate of the leaching solution in C2 and the mass of spent catalyst contained in
pared as described above (with the culture medium, powdered C2 for runs #1–4.
sulphur and the spent catalyst at different pulp densities), but the
inoculation (with washed cells) was made after 24 h, prior to a
new adjustment of the pH through the addition of sterile 1/10 2.4. Precipitation experiments
sulphuric acid (for that purpose, about 0.7–1 ml was needed).
To evaluate if A. thiooxidans was able to leach the nickel Cultures of Desulfovibrio sp. were obtained by filtering a
from the spent catalysts without adding sulphur, a flask with 1% grown culture (2 × 108 bacteria/ml) in Postgate’s B with blue
pulp density was inoculated with unwashed cells. Control exper- ribbon paper and adding this to 10 ml of Postgate’s C medium
iments were conducted using sterile culture medium. Cultures (10%, v/v) [18]. Anaerobic conditions were achieved by adding
were incubated at 30 ◦ C in a rotary shaker at 150 rpm. All exper- thioglycolate and ascorbic acid (0.1 g/l each) to the samples.
iments were carried out in duplicate. Samples were withdrawn at Different volumes of the nickel leachates from the column C2
regular intervals for the analysis of pH, nickel concentration and (see Section 2.3.3) were added to the cultures to give final
bacterial population. Samples were taken the following days: 1st, nickel concentrations in the range of 18–180 ppm. Samples were
2nd, 8th, 15th and 22nd. Evaporation was compensated prior to withdrawn after 1 week for analysis of the nickel and sulphate
sampling by the addition of sterile distilled water. concentrations.
A bioleaching experiment in a 0.85 l stirred tank reactor was Indirect precipitation experiments using sulphide biogener-
also carried out. The reactor contained 0.75 l of iron-free 9K ated by Desulfovibrio cells were also performed. In this case,
medium, 7.5 g of powdered elemental sulphur, 10 g of spent batch cultures of Desulfovibrio sp. in Postgate’s C medium were
catalyst and 75 ml of inoculum (taken directly from a grown cul- prepared in 250-ml Erlenmeyer flasks completely filled with
ture), the mixture was magnetically stirred and the temperature liquid and sealed with a rubber stopper. Cultures were run in
maintained at 30 ◦ C. batch mode until further reduction of sulphate was not detected.
Afterwards, the exhausted medium containing the biogenerated
2.3.3. Bioleaching using sulphuric acid generated by an A. sulphide was pumped at different rates into a column containing
thiooxidans biofilm the leachates. The volume of leachate and the inlet rate were
Experiments were carried out using two percolator columns. changed in the different runs. The concentration of sulphide in
In the first percolator column (C1), A. thiooxidans was immo- the inlet solution and the final concentration of soluble nickel
bilised on elemental sulphur particles to produce sulphuric acid. were measured.
The column contained 145 g of sulphur (particle size 2–4 mm),
230 ml of culture medium and 20 ml of inoculum. The immobil- 2.5. Analytical methods
isation was achieved by recirculating the medium in the column
until the pH was around 0.8. At that moment, the exhausted Dissolved sulphate was determined by a turbidimetric
medium was replaced by fresh medium (without inoculation) method [19]. Sulphate forms an insoluble precipitate with bar-
and the procedure was repeated three times to ensure that the acid ium (BaCl2 ) under acidic conditions. The absorbance of the
was produced by the attached cells. After the third replacement, sample was measured at 450 nm using a Beckman DU 640
once the pH of the culture reached a value of approximately Spectrophotometer (Fullerton, CA, USA). Samples were cen-
0.8, the acidic medium was collected and fresh medium was trifuged for 2 min at 14,000 rpm and diluted before performing
added to C1. The collected medium was pumped at a rate of the analysis. Dissolved sulphide generated in Desulfovibrio sp.
113 ml/h into the second column (C2), which contained the spent cultures was measured by titration using standard iodine and
catalyst. sodium thiosulphate solutions and starch as indicator (iodo-
It was observed that the simple percolation of the leaching metric method) [19]. Dissolved nickel in the leachates and
medium through the spent catalyst did not produce a signifi- in the digested catalyst was determined by atomic absorption
cant solubilisation of nickel, because of this, the medium was spectrophotometry (Shimadzu AA6650, Shimadzu Corporation,
forced to recirculate in C2. The amount of spent catalyst as Kyoto, Japan). A standard solution of 1000 mg/l of nickel (Carlo
well as the recirculation rate in C2 were changed in different Erba, Milan, Italy) was used to prepare the calibration stan-
runs. The conditions employed in each run are summarized in dards. Samples were diluted with 0.14 M HNO3 and filtered
Table 1. through a 0.45 ␮m membrane. The sulphuric acid produced by

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bacterial oxidation of sulphur was analysed by titration with increase in pH and this inhibited the bacterial growth, especially
0.01 M NaOH. Free bacterial population was determined by in the cases of the higher pulp densities. It was observed that
counting in a Petroff Huaser chamber in conjunction with an after 1 week the pH rose from 5.6 to 7.1 (in the case of 1%
optical microscope with phase contrast (Nikon Labophot, Kyoto, pulp density) and 7.2 (in the case of 2% pulp density) and the
Japan). number of bacteria decreased from the initial level of 5 × 107 to
2 × 107 bacteria/ml. Negligible nickel dissolution was observed
2.6. Statistical analysis in these cultures.
Two different strategies were investigated in an effort to
All experiments were conducted in duplicate. Nickel con- reduce the inhibition resulting from the pH increase. Firstly,
centration was measured three times on each duplicate sample. cells were inoculated from a grown culture without washing
Data points in figures represent means with error bars shown (unwashed cells); in this case, the acidity of the inoculum
(± S.D.). Both one-way and two-way analysis of variance were avoided the rapid increase in pH and the consequent inhibition of
performed where appropriate (with 95% level of confidence, bacterial growth. However, only in the lowest pulp density was
α = 0.05), using the statistical package on Microsoft® Office there a decrease in pH, an observation that suggests adequate
Excel Version 2003. Also, t-test was used to compare leaching bacterial growth. The second strategy involved postponing the
efficiencies. inoculation (with washed cells) for 24 h prior to a new adjust-
ment of the pH through the addition of sterile 1/10 sulphuric
3. Results and discussion acid (about 0.7–1 ml was needed).
The percentages of nickel leached using these last two strate-
3.1. Leaching experiments gies for the different pulp densities are shown in Fig. 1. It can be
seen from the figure that systems inoculated with washed bacte-
3.1.1. Chemical leaching experiments in Erlenmeyer flasks ria were less efficient than those inoculated with unwashed cells
Treatment of 1 g of the spent catalyst with diluted sulphuric for the highest pulp densities. Two-way ANOVA, performed
acid solutions gave, after 8 days, good leaching efficiencies considering the pulp density and the inoculation mode as the
in systems with initial pH values of 1 or 2 (100 and 73%, two independent factors, indicated that there was a significant
respectively, S.D. < 1%). Significant dissolution of nickel was difference between the leaching percentages obtained. t-Test
not observed in samples with pH values higher than 3 (32 and performed at each pulp density also showed that there were sig-
10% for pH values 3 and 5, respectively; S.D. < 1%). In all cases nificant differences in the leaching efficiencies obtained with
an increase in the pH was observed. Analysis of variance con- each inoculation procedure.
sidering one factor (the pH of the solution) indicated that there It was found that the higher the pulp density the lower percent-
are significant differences between the leaching efficiencies of age of dissolved nickel. This result is unexpected because the
the different solutions for a level of confidence of 95%. amount of sulphur employed would produce sufficient sulphuric
Typically, spent nickel catalysts contain metallic nickel and to react with all of the nickel present in the spent catalyst. Similar
nickel oxide, although nickel sulphides may occasionally be
present along with mixtures of coke, hydrocarbons or fat [20].
Thermodynamic data (Pourbaix Diagram) shows that Ni0 can be
spontaneously oxidised to Ni2+ at pH values lower than 4.3–4.4
according to the following equation:

Ni(s) 0 + 2H+ (aq) → Ni2+ + 2H2 (g) (1)

while the dissolution of nickel hydroxide can be achieved at pH

values below 6.5 [21]. Both processes – the oxidation of Ni0 and
the dissolution of nickel hydroxides/oxides – led to an increase
in the pH of the solution. The low-level of nickel dissolution
in the chemical leaching at pH 5 suggests that there is a small
amount of oxidised nickel present in the spent catalyst. A simi-
lar trend in nickel dissolution behaviour with pH was observed
by Cerruti et al. [22] in a study of spent nickel–cadmium
batteries as opposed to spent catalyst. Al-Mansi and Abel
Monem [23] reported similar results for the recovery of
nickel from a spent catalyst, albeit with a much higher acid
concentration. Fig. 1. Percentages of leached nickel in the bioleaching experiments in Erlen-
meyer flasks using different pulp densities. Data represent the average of two
independent samples analysed in triplicate, bars represent the standard devia-
3.1.2. Bioleaching experiments tion. Two-way ANOVA test with two factors showed that all the results have
When the bioleaching experiments were carried out using significant differences (α = 0.05). According to the t-test performed at each pulp
washed bacteria, the alkalinity of the spent catalyst produced an density, the results have significant differences (α = 0.05).

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behaviour was observed by Cerruti et al. [22] on using A. ferroox-

idans for the leaching of Ni–Cd batteries. They concluded that
there is competition between the two surfaces for the attachment
of bacteria on increasing the pulp density; thus, at the higher pulp
density a large number of bacteria are attached to the spent cata-
lyst and, consequently, they are unable to grow (initial bacterial
attachment to the sulphur is required for its oxidation) and a
smaller amount of sulphuric acid is produced. A similar effect
of pulp density was observed in the leaching of nickel, molyb-
denum and aluminium from a spent refinery processing catalyst
using adapted Aspergillus niger cultures [24]. According to the
authors, the decrease in the level of extraction of the three metals
was due to the toxicity of heavy metals. However, acidophilic
microorganisms are able to inhabit metal-rich environments
(such as acid mine drainage, for instance). These microorgan-
isms are very tolerant to a wide range of metal ions. The degree Fig. 2. Percentages of leached nickel obtained in the different runs using the
acidic medium generated by the A. thiooxidans biofilm (see conditions of each
of metal resistance varies according to the strain and the culture run in Table 1). Data represent the average of three replicates, bars represent the
conditions, but in general it is at least one order of magnitude standard deviation.
higher than the tolerance exhibit by neutrophilic microorganisms
[25]. It has been reported that A. ferrooxidans was able to growth 3.1.3. Bioleaching using sulphuric acid generated by an A.
in the presence of 30.0 g Ni(II)/l [26] while adapted strains have thiooxidans biofilm
been isolated that can grow in the presence of up to 58.7 g/l [27]. Sulphuric acid generated by A. thiooxidans growing on ele-
Unfortunately, there are very few reports concerning the toxicity mental sulphur particles in column C1 was used as leaching
of heavy metals to A. thiooxidans. Although it has been reported medium. The culture medium pH ranging from 0.52 to 0.8 was
that sulphur oxidation by A. thiooxidans was strongly inhibited pumped at rate of 113 ml/h to C2. The volumes pumped var-
by 0.30 g Ni(II)/l [28], preliminary experiments performed in ied between 140 and 200 ml. The volumes and the pH of each
our laboratory with the strain of A. thiooxidans used through- leaching medium used are shown in Table 1. As it was indi-
out the present work showed there was not growth inhibition cated in Section 2.3.3, the simple percolation of the leaching
(measured as the decrease of pH culture) in the presence of sol- medium through the spent catalyst did not produce a significant
uble nickel up to a concentration of 2.0 g/l (data not shown). That solubilisation of nickel. The leaching of nickel increased when
concentration is much higher than the maximum achieved in our the leaching solution was forced to recirculate through the spent
experiments. catalyst. The pulp density was increased from an initial value of
The percentage of nickel leached by the sterile control 10–15 g which was the maximum possible without flooding. In
(medium 0K, pH 5) was higher than that obtained in the sul- these conditions, an increase in the recovery of nickel was found.
phuric acid solution of similar pH. The presence of NH4 + in the Increasing the recirculation rate produced a decrease in the per-
culture medium at that pH value could enhance the dissolution of centage of leached nickel. The percentages of nickel leached
nickel through the formation of the complex Ni(NH3 )6 2+ , which for each run after 24 h of recirculation are shown in Fig. 2. It
is more stable than Ni(H2 O)6 2+ [29]. can be seen from the figure that the recirculation rate was the
In order to analyse whether the bacteria could grow in the most important factor affecting the leaching efficiency. When
presence of the spent catalyst without the addition of sulphur, the mass of the residue was increased by 50% (runs 1 and 2),
an experiment was performed in which 1% pulp density (and no while the inlet rate of the leaching medium and the recirculation
sulphur) was added to the culture medium. In this case, there was rate were constant, the percentage of leached nickel increased
an initial dissolution of nickel due to the acidity of the inoculum from 71 to 92%. When 15 g of spent catalyst were leached at the
but this did not increase even after 30 days. same inlet rate but with higher recirculation rates (runs 3 and 4),
The possibility of scaling up the process was investigated by the efficiency of the leaching process diminished (54 and 49%,
performing bioleaching experiments in a 0.85 l stirred tank reac- respectively)—almost certainly due to incomplete contact.
tor. The reactor contained 0.75 l of iron-free 9K medium, 7.5 g
of sulphur, 10 g of spent catalyst and 75 ml of inoculum (taken 3.2. Precipitation experiments
directly from a grown culture) and the mixture was magnetically
stirred with the temperature maintained at 30 ◦ C. Approximately Experiments were carried out in sealed flasks by adding dif-
340 mg of nickel were leached from the waste in 48 h, represent- ferent volumes of the leaching nickel solution to Desulfovibrio
ing 80% of the total nickel in the reactor. Although the recovery sp. cultures. This did not allow the development of the bacteria
of nickel was significant, the system was not particularly good or, therefore, the production of sulphide. Sulphate reducing bac-
from a practical point of view. The physical characteristics of the teria are sensitive to the presence of heavy metals. Experiments
spent catalyst (a very fine powder) made it adhere to the reac- involving nickel precipitation performed in our laboratory
tor wall, resulting in an inhomogeneous system with a lower showed that Desulfovibrio sp. could grow in the presence of up
efficiency. to 25 ppm of dissolved nickel, but the results were highly depen-

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Table 2
Conditions employed and results obtained in the precipitation experiments using biogenerated sulphide
Q (ml/min) m(S2− ) (mg) m(Ni)i (mg) m(Ni)f (mg) Percentage of Ni precipitated

A 5 22.74 68.37 14.3 79

B 5 27.01 136.7 96.6 29
C 5 33.47 205.1 201.2 2
D 10 29.29 136.7 113.3 17

Q: inlet flow rate of the solution containing biogenerated sulphide; m(S2− ): mass of dissolved sulphide generated; m(Ni)i : mass of nickel initially present in the
precipitation column; m(Ni)f : mass of soluble nickel after the contact with the sulphide solution; percentage of Ni precipitated: (m(Ni)i − m(Ni)f ) × 100/m(Ni)i .

dent of the inoculum conditions. According to data reported in sulphides are more insoluble than phosphates and oxalates; in
the literature, nickel concentrations between 10 and 20 ppm are addition the culture of SRB does not require complex organic
toxic for pure and mixed cultures of SRB [30]. To avoid the inhi- compounds as Citrobacter does [9].
bition produce by the presence of nickel on the bacterial growth, Nickel could have been recovered from the leaching solution
the precipitation was carried out indirectly. Thus, the recovery as solid NiSO4 by evaporation for 5 h at 100 ◦ C as proposed
of nickel from the leaching solution was performed using by several authors [8,23,34]. However, the solubility of NiS is
sulphide generated in a Desulfovibrio sp. spent culture. The much lower than that of the NiSO4 (0.00036 and 29.3 g per
results obtained in each experiment are summarized in Table 2. 100 ml for NiS and NiSO4 , respectively), thus allowing the
It can be seen that the highest precipitation efficiency (79%) is recovery of nickel as a solid salt at room temperature. Beside this,
achieved for the lowest inlet rate and the lowest mass of nickel. the low-solubility of nickel sulphide would allow a safer final
That percentage of nickel precipitation represents the maximum disposal.
possible value according to the amount of sulphide introduced
into the system. When the mass of soluble nickel was increased 4. Conclusions
(experiment B) only 29% of the nickel was precipitated and this
corresponds to 81% of the sulphide in the inlet. Thus, 19% of The results of our experiments show that nickel present in
the sulphide entering the system did not react with the dissolved the spent material from the hydrogenation of vegetable oil can
nickel, probably because a proportion of the sulphide was lost be successfully leached using A. thiooxidans. The percentages
as H2 S produced during contact with the acidic leachate. of nickel leached using A. thiooxidans where higher than those
A process for the indirect precipitation of heavy metals by obtained with dilute sulphuric acid solutions. Batch experiments
sulphate reducing bacteria has been previously studied [12]. This showed a decrease in the leaching efficiency on increasing the
method involves the production of H2 S by SRB in a fixed-bed different pulp densities—probably due to a competition between
column reactor. A gas mixture of H2 and CO2 was injected into the spent catalyst and the sulphur for bacterial attachment. The
the column while the SH2 was removed from the outlet gas physical characteristics of the residue meant that the best results
flow by N2 and it was bubbled into a reactor containing a heavy were obtained when the leaching process was carried out using
metal solution. Although the system allowed copper and zinc sulphuric acid biogenerated by an A. thiooxidans biofilm. The
to be selectively recovered, a preliminary estimation of costs recovery of nickel from the leachates was performed at room
indicated that, in that particular case the treatment would not temperature by precipitating with sulphide generated by Desul-
be economically viable. A simple and low-cost system for the fovibrio cells. Due to the toxic effects of nickel ions on the growth
treatment of acid mine drainage has been designed. The sys- of Desulfovibrio, direct precipitation was not possible. Indirect
tem consisted in a biofilm reactor inoculated with acidophilic precipitation using sulphide generated in Desulfovibrio sp. cul-
SRB immobilised on porous glass beads [31]. Although these tures allowed the recovery of nickel as the very insoluble nickel
SRB populations were able to efficiently reduced sulphate to sulphide. In order to improve the efficiency of this process, the
sulphide, it is necessary to consider that this type of reactor, possibility of sulphide volatilisation should be minimised.
where a heavy metal solution percolates the biofilm is subject
to clogging because of the accumulation of insoluble metal sul- Acknowledgements
phides. The generation of sulphide in a simple reactor as a batch
reactor can be used in the precipitation of heavy metals without Dr. Edgardo Donati and Dr. Marisa Viera are research mem-
the design of more complex reactors or the needing of gas car- bers of CONICET. This research was supported in part by
riers, which increase the operational costs. Although the use of ANPCyT (PICT 25300) and CONICET (PIP 5147).
SRB to precipitate heavy metals has been widely studied, this
is no the only possibility for metal precipitation. Precipitates of References
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Please cite this article in press as: V. Bosio, et al., Integrated bacterial process for the treatment of a spent nickel catalyst, J. Hazard. Mater.
(2007), doi:10.1016/j.jhazmat.2007.10.095
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Please cite this article in press as: V. Bosio, et al., Integrated bacterial process for the treatment of a spent nickel catalyst, J. Hazard. Mater.
(2007), doi:10.1016/j.jhazmat.2007.10.095