Biological Beneficiation of Iron-Ores PDF

Delvasto, P., Ballester, A., Muñoz, J.A., González, F., Blázquez, M. L., García-Balboa, C., 2005.
Exploring the possibilities of

biological beneficiation of iron-ores: The phosphorus problem. In: Proceedings of the 15th Steelmaking Conference, 5th
Ironmaking Conference & 1st Environment and Recycling Symposium IAS (CD-ROM). Argentinean Steelmaking Institute (IAS).
San Nicolás, Buenos Aires, Argentina, November 7-10, 2005. pp 71-82
Exploring the possibilities of biological beneficiation of iron-

ores: The phosphorus problem
P. Delvasto1, A. Ballester1, J.A. Muñoz1, F. González1, M.L. Blázquez1, C. García-Balboa2
1
Biohidrometallurgy Research Group. Department of Materials Science and Metallurgical Engineering.
Complutense University. 28040 Madrid. Spain.
2
Department of Industrial Technology. Alfonso X “el Sabio” University. Villanueva de la Cañada, Madrid, Spain
Synopsis
In past decades, preferences of international iron-ore trade markets have moved to low-
phosphorus iron ores because of the well-known detrimental effects of phosphorus on
steel and other iron-related materials [1, 2]. This fact has made that in most places only
low-phosphorus ore is exploited making premium grade resources more scarce and
leaving mines enriched in high-phosphorus ore [2]. Although it is a well established fact
that these type of ores can be treated by roasting and acid-leaching processes [2, 3],
traditionally low prices of iron ores make these processes not attractive from the
economical standpoint. Moreover, such processes imply practices that, if not carefully
considered, can be potentially deleterious for the environment. Biotechnology can bring a
solution to overcome the afore mentioned problems. Since phosphorus is a major nutrient
for most living forms, biological extraction of phosphorus from iron ores can become
reality. Little attention has been paid to biological dephosphorization or iron ores. Some
works [4, 5] have dealt with the use of fungal metabolites to attain dephosphorization,
however not very high yields have been reported. More recently, phosphorus-containing
blast furnace slag has been successfully leached using heterotrophic bacteria [6]. The
main drawback of these investigations relies on the fact that species used for
beneficiation were not related to the ore being treated. In biological beneficiation
processes it is always recommendable to employ indigenous species to reduce
environmental impacts.
In this work, authors report the successful activation of the indigenous microflora
associated to a rejected high-phosphorus iron ore from a Brazilian mine. By using the
shake-flask technique, samples of the ore were amended with glucose-containing liquid
medium to promote the growth of microorganisms naturally present in the ore. It was
found that microflora was complex and in some cases its growth was led by fungi while
in others biomass was predominantly composed by bacteria. The microflora was screened
to detect species able to solubilize phosphorus using standard microbiological techniques.
In such a way bacterial species of genus Burkholderia and fungus of genus Aspergillus
showing phosphorus solubilization were isolated from ore microflora. It was found that
in bacterial-led experiments dephosphorization attained was around 10% while in fungi-
led experiments dephosphorization of the ore reached 20%. The results are discussed and
an analysis of the use of the shake-flask technique in this research is criticized. Fore-
coming research is also outlined.
71
Delvasto, P., Ballester, A., Muñoz, J.A., González, F., Blázquez, M. L., García-Balboa, C., 2005. Exploring the possibilities of
1. Literature review
Two major routes have been traditionally proposed for the dephosphorization of iron
ores: physical and hydrometallurgical routes [1]. Choosing one or another will strongly
depend on the characteristics of the ore as well as the degree and type of association
between iron minerals and phosphorus. When phosphorus is present as a product of
primary mineralization, i.e. occluded phosphates in an iron oxide matrix,
dephosphorization can be attained through a combination of physical processes such as
mechanical liberation and flotation [1]. However this is not always the case. Some Iron
ores are composed by secondary minerals produced through weathering of primary rocks.
This type of ores exhibit such an intimate relationship between P and Fe-oxides that it
does not result easy to elucidate the real nature and degree of association between both
phases. For this type of ores, the hydrometallurgical route has been the usual approach [1,
2, 9]. Acid or alkaline leaching of previously roasted high phosphorus iron ores has been
described in several research papers as well as patents, as shown on table 1. As seen on
table 1, dephosphorization yields using hydrometallurgical routes range between 60%
and 97%.
Table 1. Summary of conditions for several chemical treatments for

dephosphorization of iron ores.
Maximum
Ore Roasting Roasting
Leaching agent removal of Reference
composition conditions additives
P (%)
Iron oxide 900 ºC
LiCl Sulfuric Acid 96.4 [7]
(unspecified ) 2 hr.
Iron oxide 900 ºC
CaCl2 Nitric Acid 96.6 [7]
(unspecified ) 2 hr.
Iron oxide 900 ºC Hydrochloric
BaCl2 85.7 [7]
(unspecified ) 2 hr. Acid
Hematite 1200 ºC
none Sulfuric Acid 68.0 [8]
Goethite. 1 hr.
Hematite 1100 ºC Sodium
none 63.0 [8]
Goethite 1 hr. hydroxide
Magnetite none none Nitric Acid 75.0 [3]
Magnetite
1200 ºC Hydrofluoric
Hematite none 72.7 [9]
1 hr. Acid
Goethite
Magnetite
1200 ºC Hydrochloric
Hematite none 61.8 [9]
1 hr. Acid
Goethite
Hematite 1250 ºC
none Sulfuric Acid 68.7 [2]
Goethite hours
Although this kind of processes have demonstrated high efficiency, they involve energy
consuming steps as well as handling of highly dangerous substances. In addition, iron ore
is a commodity of traditional low price, so the cost of additional facilities for
72
dephosphorization in iron ore processing plants is not always justified, neither from the
economical nor the environmental standpoint. This fact has made researchers to turn their
attention to biological dephosphorization processes.
P is a limiting nutrient for all living forms. It is a well documented fact that, under
starvation conditions, microorganisms can utilize phosphorus from mineral sources such
as feldspars and phosphatic rocks [10]. In fact, it has been determined that phosphorus
containing minerals can be biologically solubilized easier than non-phosphorus-
containing minerals [10]. This ability has been extensively investigated and applied in
soil science and agriculture [11], because of its implications on the fertility of soils. The
accumulation of these evidences reveals that microbiological activity can be useful to
remove phosphorus from iron ores. Despite of it, few works can be found in the literature
regarding to this field.
An interesting work by Parks et al. [5] characterized all metabolic products generated by
a fungus, Penicillium sp., while growing in contact with a high phosphorus iron ore
concentrate. These researchers found that this fungus was able to produce itaconic and
oxalic acids when consuming glucose as carbon source. The dephosphorization degree
reported in this work reached 20%. They also tried mixtures of fungal metabolic products
with HCl, finding that dephosphorization could be increased up to 50%. Buis [4], studied
the dephosphorization capacity of several fungi including: Paxillus involutus, Hebeloma
crustiniforme, Thelephora terrestris and Laccaria bicolor. Although the fungi studied by
this researcher were able to solubilize hidroxylapatite, a heavily insoluble phosporus
compound, the solubilization of phosphorus from iron ores was so poor, reaching around
1%. More recently, some researchers from India [6] have used a soil bacterium,
Frateuria aurentia, to solubilize phosphorus from a LD slag (30%Ca, 20%Fe, 5% Mg,
1.4%P), a steelmaking byproduct. They report that this bacterium can solubilize between
72% and 90% of the P present in this iron slag, making this material able to be re-
circulated, as fluxing material, into the blast furnace operations.
2. Materials and methods
2.1 Sample characterization
The high-P iron ore employed throughout the study was the rejected fraction from a
magnetic separation process. The origin of the ore is the “Jangada” mine in the region of
Minas Gerais, Brazil. This ore underwent a crushing process so its size ranged between
7.0 mm and 0.03 mm with an average size of 2.3 mm. X-ray diffraction analyses revealed
hematite and quartz as main mineralogical species present in the ore, with some goethite
also present. X-ray fluorescence elemental chemical analysis is shown on table 2.
73
Table 2. Main components (w%) of the iron ore, as obtained by elemental X-ray
fluorescence chemical analysis.
Fe O Si Al Mn P Cr Ti
55,27 33,30 9,21 1,58 0,24 0,26 0,059 0,038
2.2 Isolation of the microorganisms present in the ore
Since this work is focused on the phosphorus problem, microbiological techniques were
used to screen microorganisms naturally present in the ore exhibiting phosphate
solubilization capacity. The problem was approached applying procedures usually
described for the isolation of phosphate solubilizing microorganisms (hereafter PSM)
from soils [11]. Cells and spores were detached from ore surface by shaking at 150 rpm a
mixture of 5 g of fresh mineral and 100 ml of sterile distilled water for 24 hours.
Supernatant was serial diluted in sterile distilled water and suitable aliquots spread over
Petri dishes containing a differential growth media for PSM. The growth media
composition is shown on table 3.
Table 3. Composition (per liter of distilled water) of the PSM differential media employed in this study.
Component gr/l
Glucose 10.0
Ca3(PO4)2 2.5
Agar-Agar 20.0
Nutrient salts
50.0
solution (ml)*
*Per liter: MgCl2·6H2O, 100 g; MgSO4·7H2O, 5 g; KCl, 4 g; (NH4)SO4, 2 g.
This isolation technique is a standard plate assay method for isolation of PSM based on
the formation of a solubilized halo around the colonies of microorganisms capable to
solubilize a calcium phosphate insoluble compound [12]. Inoculated plates were
incubated at 30 ºC for at least 12 days. To obtain pure PSM strains, colonies were
collected using a loop and streaked upon fresh medium plates. The isolates were then sent
to specialized laboratories for its characterization by molecular –genetic techniques.
2.3 Shake-flask leaching experiments
Two types of shake-flasks leaching tests were carried out: Bioactivation of the ore by
supplying a nutrient solution directly to the fresh ore and leaching of the previously
sterilized ore using a pure PSM isolate.
74
In the first case 10 g of fresh ore with a particle size of 2 mm was added under sterile
conditions into Pyrex bottles containing 100 ml of sterile glucose-based nutrient broth
(per liter: glucose 10 g and 50 ml of nutrient salts solution). Up to 18 replicates of this
experiment were performed and run over 70 days. pH of the broth was regularly
measured. At the end of the experiment, the mineral was washed with a 5% sodium
hypochlorite solution under shaking to separate the biomass, washed repeatedly with
distilled water and pulverized in a ball mill. Remaining P content was determined by X-
ray fluorescence spectroscopy (XRF).
In the second case, 0.2 mm ore was sterilized by autoclaving to prevent interference with
other microorganisms present in the ore. Using the same procedure described above, 7.5
g of the ore was added to 150 ml of glucose nutrient broth. Then the reactor was
inoculated with 0.2 ml of an Aspergillus niger spore suspension. This fungus was isolated
from the ore, as shown on section 3.1. The process was monitored by measuring pH as
well as iron content of the leaching liquor, determined by atomic absorption
spectroscopy. At the end of experiments the ore was treated similarly as for the
bioactivation experiments and the final P content in the ore was determined using XRF as
well. In this case all experiments were prepared by triplicate.
3. Results and discussion
3.1 Bioactivation of ore microflora
As a first approach, the microflora naturally associated to the ore was activated by means
of the addition of a phosphorus-free glucose-based nutrient liquid. The aim of this
experiment was to force microflora to utilize the phosphorus present in the ore. The
experiment revealed the complexity of the ore microflora. Several repetitions of this
experiment were performed, and it was found that in 31% of the cases bacterial
populations dominated exclusively the microcosms, 56% of the microcosms were
dominated exclusively by fungi and 13% of the microcosms exhibited a mixed behavior.
Figure 1 (a) depicts the typical in-flask and in-plate features of a microcosm dominated
by fungi while figure 1(b) shows the same in the case of bacterial domination.
Bacteria-dominated microcosms became turbid after 12 days of culturing. Fungi-

dominated microcosms, on the other hand, generated clear supernatants and abundant
fungal filaments growing intermixed with the ore particles. These two types of behavior
also showed physiochemical differences, as depicted in figure 2, where the pH evolution
throughout the entire experiment is shown. It can be seen that within the first 5 days of
the experiment both kind of microcosms have a similar behavior. After this time,
bacterial-led microcosms increased the pH values above the initial pH of the system,
reaching a maximum at day 12, coinciding with the beginning of the turbidity in the
supernatant. Fungi-led experiments exhibited a pH drop until day 12, for later increase
softly but always below the initial pH value. At the end of the experiment, day 70th, the
ore was separated and analyzed for P.
75
Figure 1. Bioactivation of the iron ore with a glucose-based medium. The complexity of the ore
microflora led to two main types of microbiological behaviors. In (a) Fungi as predominant species and
(b) Bacteria as predominant species.
7.00
6.00
5.00
Average pH
4.00
3.00
Bacteria
2.00 Fungi
1.00
0 20 40 60 80
Time, days
Figure 2. Evolution of pH in bioactivation experiments. The difference exhibited between bacterial led
experiments and fungi led experiments is shown.
76
The percentage of dephosphorization attained in the case of bacteria-led experiments was

10% while in the case of fungi-led experiments increased up to 20%. This evidences the
importance of acidification for the dephosphorization process. Despite it was not
measured, visual inspection of the cultures indicates that produced fungal biomass was
higher than bacterial biomass. Since P is a structural component of living cells, it can be
hypothesized that fungi-led microcosms can incorporate more of the mobilized P. To
elucidate how this complex microflora was composed and select those microorganisms
with the higher capacity to solubilize mineral phosphates, a microbiological screening
procedure was performed.
3.2 Isolation of phosphate solubilizing microorganisms from the ore
Phosphate solubilizing microorganisms (PSM) is a generic classification that includes

several bacteria and fungi that are able to mobilize phosphorus from hardly soluble
phosphorus sources, like mineral phosphates. Using a standard technique, based on the
in-plate solubilization of calcium phosphate, it was found that the iron ore analyzed in
this work had up to five PSM species among its indigenous microflora. The identified
species are shown on table 4. In figure 3, the halo formation is depicted for three of the
isolates.
Table 4. PSM isolated from the iron ore microflora
Identified
Type Halo generation
species
Bacterium Burkholderia +
sp.
Bacterium Clavibacter xyli +/-
caribiensis
cepacia
Filamentous fungus Aspergillus +
niger
It must be pointed out that several other unidentified species, especially fungi (4 more)
were isolated, however were not selected because of their poor or inexistent
solubilization ability. It has been reported [13] that Burkholderia species produce some
anti-fungal compounds. This could explain why in bacteria-led microcosms fungi are not
present. On the other hand, if fungi are able to develop faster, the acidification of the
liquid media, due to fungal growth, will suppress bacterial growth. An intermediate
growth condition would explain the mixed behavior exhibited by 13% of the microcosms.
Experiments made in liquid medium (data not shown) showed that the best phosphorus
solubilizer was the isolated fungal strain Aspergillus niger. This isolate was chosen for a
more complete experiment of dephosphorization of the ore.
77
Figure 3. Phosphate solubilization halo (dark zones) in plate cultures of some PSM isolated from iron ore.
(a) Bacterial colonies of Burkholderia sp. and (b) Burkholderia caribiensis. (c) Fungal colonies of
Aspergillus niger. Scale bar: 1 mm.
3.3 Dephosphorization of the ore using isolate Aspergillus niger
Due to the complexity derived from the multiple interactions between the different
species present in the ore microflora, it was decided to investigate the iron ore
dephosphorization using a pure isolate. In this case Aspergillus niger was chosen. It is
well known the ability of this microorganism for generating different leaching substances
including citric, oxalic and gluconic acid [14]. This property has been successfully used
in heterotrophic leaching and beneficiation of different minerals including Ni and Co
laterites, clays, Cu minerals and silicates [14]. In figure 4 can be observed that A. niger is
able to mobilize slightly more than 30% of the phosphorus originally present in the ore in
21 days of treatment when using glucose as carbon source. Since there is no other P
source in the nutrient solution, A. niger is obligated to scavenge the P present in the ore to
continue growing. That explains the semi-sigmoidal trend exhibited by the
dephosphorization percentage as a function of time. This is in agreement with the
evolution of pH, figure 5, where the initial value of 5.6 rapidly decreases to a value close
to 3 for then getting stabilized around 2.6.
These pH values are somehow lower than those reached in the fungi-led bioactivation
experiments, figure 2. This might be due to the competition established among all the
fungal species present in the ore microflora. If a extremely effective acid-producing
species like A. niger have a sharing-free access to nutrients, its metabolism will be evolve
into an optimum condition that permits a higher acidification of the surrounding media,
and as a consequence, a higher mobilization and uptake of the P occluded into the ore.
An important factor that should be addressed is the leaching of the ore itself. This factor
is quantified by the iron that goes into solution, expressed as percentage of iron losses in
78
figure 5. In a hypothetical biobeneficiation process of iron ore, what is important is to

leach away the impurities present in the ore, P in this particular case, with a minimum
loss of the metal value of the ore i.e. iron.
35
([ % P initial - % P final] / % P initial)* 100
30
Dephosphorization efficiency
25
20
15
10
0
0 5 10 15 20 25
Time, days
Figure 4. Dephosphorization of iron ore (particle size 0.2 mm; initial P content 0.26 w%) with an isolate
of Aspergillus niger using the shake flask technique. Pulp density 5%. The growth medium used was a
phosphorus-free, glucose-based nutrient solution.
9.00 1.000
8.00 0.900
0.800
7.00
0.700
6.00
Iron losses, w%
Average pH
0.600
5.00
0.500
4.00
0.400
3.00
0.300
2.00
0.200
Average pH
1.00 0.100
Iron losses, w%
0.00 0.000
0 5 10 15 20 25
Time, days
Figure 5. Evolution of pH and iron losses during dephosphorization of iron ore (particle size 0.2 mm
initial P content 0.26 w%) with an isolate of Aspergillus niger using the shake flask technique. Pulp
density 5%. The growth medium used was a phosphorus-free, glucose-based nutrient solution.
79
It is seen that iron losses increase as the time proceeds. Moreover, as iron losses increase,
dephosphorization increases as well. This is a natural consequence of the fungal growth
and its strategy to scavenge such an important and limiting nutrient as P is. The fungal
colonization of the ore can be observed in the scanning electron microscope (SEM)
image shown in figure 6. Fungal filaments, made up of fungal cells, colonize the ore
surface and producing organic acids which in turn dissolve those ore phases on which
phosphorus is present and not readily available to sustain fungal growth. Once these
phases are attacked, P is liberated and then utilized by the fungal cells. This ore attack is
indicated by the solubilization of the iron in the ore. So, to attain dephosphorization,
some of the iron associated with phosphorus, or enclosing phosphatic phases, should be
dissolved.
Figure 6. SEM image of Aspergillus niger filaments colonizing particles of iron ore.
Some works indicate that phosphorus can be present in iron ores associated with iron
oxy-hydroxide phases such as goethite [9]. Although the ore employed in this
investigation has not been characterized to that extent, goethite was found to be present in
the ore in little quantities. Since this phase is less stable, in the physicochemical sense,
when compared to hematite the main iron phase present in the ore, the dissolution of the
P-bearing goethite possibly present in the ore may explain the little iron losses found,
which in any case, were not higher than 1% during the period of time studied.
4. Concluding remarks
In this work, dephosphorization of iron ores using microorganisms has been proven to be
possible. Bioactivation of the ore revealed the complexity of the microflora associated to
the studied iron ore and conducted to two different microbiological behaviors: one
supported by bacterial growth and other supported by fungal growth. Microbiological
screening procedures were employed to detect those species capable to solubilize mineral
phosphates. Among the isolated species, one fungus of species Aspergillus niger was
80
used for dephosphorization of iron ore with promising results. A maximum

dephosphorization degree of 30% was reached. It is, however lower than chemical based
dephosphorization processes. More research should be conducted to improve the
culturing conditions in order to increase the dephosphorization percentage attained and
justify a possible industrial application of these findings.
Acknowledgements
Iron ore samples were kindly supplied by Professor Armando Corrêa de Araújo
(Universidade Federal de Minas Gerais) and Minerações Brasileiras Reunidas (MBR).
Financial support, in the form of Doctoral Scholarship, is also acknowledged by one of
the authors (P. Delvasto) from the National Found for Science, Technology and
Innovation of Venezuela (FONACIT) and the Simón Bolívar University of Venezuela.
References
[1] Kokal, H. (1990) “The origins of phosphorus in ironmaking raw materials and
methods of removal- A review” 63th annual meeting, Minnesota section, AIME. January
1990. pp 225-257.
[2] Cheng, C., Misra, V., Clough, J., Mun, R. (1999) “Dephosphorisation of western
Australian iron ore by hydrometallurgical process” Minerals Engineering, 12 No. 9. pp
1083-1092.
[3] Muhammed, M., Zhang, Y. (1989) “A hydrometallurgical process for the
dephosphorization of iron ore” Hydrometallurgy 21. pp 277-292.
[4] Buis, P. (1995) “ Bioremediation techniques for the removal of phosphorus from iron
ore” PhD Dissertation thesis. Mining engineering. Michigan Technological University.
(1995)
[5] Parks, E., Olson, G., Brinckman, F.; Baldi, F. (1990) “Characterization by HPLC of
the solubilization of phosphorus in iron ore by a fungus” J. Ind. Microbiol. 5. pp. 183-
190
[6] Pradhan, N., Das, B., Acharya, S., Kar, R., Sukla, L., Misra V.(2004) “Removal of
phosphorus from LD slag using heterotrophic bacteria”. Minerals and Metallurgical
Processing 21 No. 3. pp 149-152
[7] Feld, I., Franklin, T., Lampking, M. (1968) “Process for removing phosphorus from
iron ores” U.S. Patent No. 3,402,041. Sept. 17, 1968.
[8] Gooden, J.; Walker, W.; Allen, R. (1974) “ADEMPHOS – A chemical process for
dephosphorisation of iron ore”. National chemical engineering conference 1974. Process
industries in Australia – Impact and growth. Surfers Paradise, Queensland, Australia, July
10 to 12th, 1974. pp 38-49.
[9] Araujo, A.; Fonseca, D.; Souza, C. (1994) “Hydrometallurgical routes for the
reduction of phosphorus in iron ores”. A. Sutulov Memorial Volume. Vol. III. Chemical
metallurgy. IV Meeting of the Southern Hemisphere on Mineral Technology. Edited by I.
Wilkomirsky, M. Sánchez and C. Hecker, Universidad de Concepción. Concepción-
Chile. pp 83-92
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[10] Banfield, J., Barker, W., Welch, S., Taunton, A. (1999) “Biological impact on
mineral dissolution: Application of the lichen model to understanding mineral
weathering in the rhizosphere”. Proceedings of the national Academy of Sciences, USA.
Vol. 96, pp 3404-3411.
[11] Kucey, R.M.N. (1983) “Phosphate-solubilizing bacteria and fungi in various
cultivated and virgin Alberta soils”. Can. J. Soil Sci. 63. pp 671-678
[12] Nautiyal, C.S. (1999) “An efficient microbiological growth medium for screening
phosphate solubilizing microorganisms” FEMS microbiology letters, Vol. 170. pp 265-
270
[13] Ravnskov, S., Larsen, J., Jakobsen, I. (2002) “Phosphorus uptake of an arbuscular
mycorrhizal fungus is not effected by the biocontrol bacterium Burkholderia cepacia”
Soil Biology and Biochemistry. Vol. 34. pp 1875-1881.
[14] Jain, N., Sharma, D. (2004) “Biohydrometallurgy for nonsulfidic minerals-A
review” Gemicrobiology Journal, vol. 21. pp 134-144
82

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Delvasto, P., Ballester, A., Muñoz, J.A., González, F., Blázquez, M. L., García-Balboa, C., 2005.

Exploring the possibilities of

Exploring the possibilities of biological beneficiation of iron-

Table 1. Summary of conditions for several chemical treatments for

2. Materials and methods

2.1 Sample characterization

2.2 Isolation of the microorganisms present in the ore

*Per liter: MgCl2·6H2O, 100 g; MgSO4·7H2O, 5 g; KCl, 4 g; (NH4)SO4, 2 g.

2.3 Shake-flask leaching experiments

3. Results and discussion

3.1 Bioactivation of ore microflora

Bacteria-dominated microcosms became turbid after 12 days of culturing. Fungi-

The percentage of dephosphorization attained in the case of bacteria-led experiments was

3.2 Isolation of phosphate solubilizing microorganisms from the ore

Phosphate solubilizing microorganisms (PSM) is a generic classification that includes

Table 4. PSM isolated from the iron ore microflora

3.3 Dephosphorization of the ore using isolate Aspergillus niger

figure 5. In a hypothetical biobeneficiation process of iron ore, what is important is to

used for dephosphorization of iron ore with promising results. A maximum

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