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n18a02

CEPAS NATIVAS TOLERANTES A METALES PESADOS AISLADOS DEL

PASIVO MINERO DE LA MINA HUALILÁN, ARGENTINA

N. Hidalgo 1*, P. Fernández 3, D. Bustos2, M. Rosa 2, A. Senese1

1: Magister, Instituto de Investigaciones Mineras, FI, Universidad de Nacional de San Juan, Argentina.
2: Dr. Instituto de Ciencias Básicas, Universidad Nacional de San Juan, Argentina.
3: Dr., PROIMI-CONICET-, Argentina.
*E-mail: [email protected]

PALABRAS CLAVE RESUMEN

Hongos filamentosos La contaminación por metales pesados es actualmente uno de los
Cepas nativas problemas ambientales más serios y complejos de tratar. Las pilas de
Pilas de lixiviación lixiviación de las industrias mineras se caracterizan por su elevado
Metales pesados contenido en metales disueltos, además de ser considerado un hábitat
propicio para el desarrollo de microorganismos resistentes. Entre
ellos, aquellos microorganismos autóctonos con adaptaciones
fisiológicas que le permiten tolerar elevadas concentraciones de
metales pesados, los cuales son considerados como candidatos para
procesos de biorremediación. Por esto, el objetivo de este estudio fue
aislar y seleccionar cepas de hongos filamentosos con potencialidad
para la remoción de metales pesados. Se tomaron muestras de pilas de
lixiviación de la Mina Hualilán en la provincia de San Juan
(Argentina). El aislamiento de los hongos filamentosos se llevó a cabo
en medio Sabouraud-glucosa-agar con antibióticos y se identificaron
por técnicas de biología molecular (amplificación y secuenciación del
fragmento ADNr 18S y 26S, ITS1, ADNr 5.8S e ITS2). Se estudió la
tolerancia a Cu (II) y Pb (II) en placas con medio Agar papa dextrosa
(PDA) con concentraciones entre 25 y 800 ppm de cada metal. Las
condiciones de incubación fueron a 28 ºC durante un período de total
desarrollo de los hongos comparado con una placa control (sin metal),
midiendo diámetros de crecimiento. Se realizó un análisis estadístico
haciendo un análisis de varianza y prueba de comparación de
promedios Duncan, con el programa estadístico Anova multifactorial
bajo un diseño experimental completamente al azar con arreglo
factorial 2x6x1, una caja de Petri como unidad experimental, y tres
repeticiones. Como resultado se aislaron tres cepas: Penicillium
simplicissimum, Fusarium sp y Penicillium funiculosum. Los hongos
identificados presentan tolerancias a distintas concentraciones de
soluciones de los metales, donde la cepa más tolerante fue Penicillium
simplicissimum.

HEAVY METALS TOLERANT NATIVE STRAINS ISOLATED FROM MINING

WASTE OF THE HUALILÁN MINE, ARGENTINA

KEYWORDS ABSTRACT
Filamentous fungi Heavy-metal contamination is one of the most serious and complex
Native strains environmental problems to be dealt with nowadays. Leaching piles
Leaching piles from the mining industry are characterized by a high content of

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Heavy metals dissolved metals. In addition, they are considered to be a suitable

habitat for the development of resistant microorganisms. Among
such microorganisms are those native ones that are physiologically
ISSN 2256-1013, No X, pp. XX-XX, 20XX. https://doi.org/XXXX/RCM.20XX.04.058
adapted to tolerate high concentrations of heavy metals and thus, are
considered to be candidates for bioremediation processes. Taking
this into consideration, the aim of this study was to isolate and select
filamentous fungal strains with the potential to remove heavy
metals. Samples of leaching piles were drawn from Hualilán Mine
in the province of San Juan (Argentina). Isolation of filamentous
fungi was carried out in a Sabouraud-glucose-agar medium with
antibiotics, and the fungi were identified by using molecular biology
techniques (amplification and sequencing of the fragment ADNr
18S y 26S, ITS1, ADNr 5.8S e ITS2). Tolerance to Cu (II) and Pb
(II) was studied in plates with Potato dextrose agar (PDA) medium
with concentrations between 25 and 800 ppm of each metal.
Incubation conditions were at 28 °C during the period of total
development of the fungi and compared with a control plate
(without metal) by measuring growth diameters. A statistical
analysis was carried out through variance analysis and Duncan
means comparison test, with the ANOVA multifactor statistical
software under a completely randomized experimental design with
2x6x1 factorial arrangement, a Petri dish as an experimental unit,
and three repetitions. As a result, three strains were isolated,
namely, Penicillium simplicissimum, Fusarium sp and Penicillium
funiculosum. The fungi identified show tolerances to different
concentrations of metal solutions, the most tolerant strain being
Penicillium simplicissimum.

1. INTRODUCTION low growth rates in plants, death of animals

and others health alterations [1].
Heavy metal contamination is one of the
most serious environmental problems During the excavation and extraction of
nowadays. Industries such as mining, metal minerals in the mining activity and,
smelting, fuel and energy production from depending on both, the characteristics of the
petroleum, the fertilizer and pesticide mining method (either underground or open-
industry and their applications, among pit), and the geological characteristics of the
others, produce waste that contains high site, there is an important movement or
concentrations of heavy metals. These displacement of land and rocks [2]. The ore
effluents are dumped into the environment extracted from the mine is subjected to a
and distributed in the soil, water and air. process of benefit and concentration that
Heavy metals are not chemically or results in the generation of a large amount of
biologically degradable like most organic waste or fragments of the mineral that
compounds; therefore their accumulation in sometimes contain a significant amount of
nature is highly dangerous. The occurrence metals or residues of the chemical used in
of heavy metals affects biological diversity the concentration. In the concentration stage
and cause serious ecological problems such of the mineral, chemical reagents, in
as the increase of bioaccumulation and the particular, cyanide and sulfuric acid are
magnification of toxins in the food chain, frequently used, the residues of which
resulting in several diseases such as lungs remain in the fragments of the mineral that is
and digestive tract irritation and/or cancer, discarded. The contact of the rain with this

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waste produces contamination of the Fungi under stress may adapt to the above
draining water [3]. conditions either through a temporary
alteration in their development patterns or
It is known that heavy metal-tolerant through changes in their physiological
microorganisms, which generally coincide characteristics, depending on the toxicity of
with those that have been isolated from metals, which in turn is influenced by the
areas contaminated by those heavy metals existing concentration of the metal [9].
[4, 5] are usually good candidates to be
studied and applied in biological removal Taking the above facts into consideration,
technologies as a sustainable alternative of this work describes the isolation of native
lower impact on the environment. Various strains of microorganisms present in waste
types of tests exist such as strength, from the Hualilán Mine of the Province of
tolerance and Minimum Inhibitory San Juan, Argentina in order to identify and
Concentration (MIC) that are used to select select those species tolerant to heavy metals.
the best strains when heavy metal dissolved
ions have to be removed. These tests are
intended to determine whether the strains 2. EXPERIMENTAL METHODOLOGY
are able to grow in an environment
contaminated with such metals. Tolerance 2.1. Sampling, chemical analysis and
tests, in turn, clearly establish a comparison visual identification of minerals of the
between the potential ability to grow in a solid
contaminated environment and the natural
ability to grow [6]. The samples were collected from 10
different points in the leach pile (waste
In the area of microbial ecology, there is an material) from the Hualilán mine, province
increased interest in the study of microbial of San Juan (Argentina), an abandoned mine
diversity and the dynamics of its where predominant elements such as gold
populations. This helps us deepen our and silver were extracted together with
knowledge both on the composition of the subordinate elements such as copper, zinc
communities existing in contaminated soils and lead (Figure 1).
and on their evolution during the pollutants
removal processes. It also helps determine Sampling was performed manually by
which microorganisms are capable of digging with shovel and approximately 500
adapting to and exploring contaminated g of sample was removed from each selected
habitats [7]. By means of molecular point. The material used was previously
techniques, the structure and composition sterilized and the samples were stored in
of the microbial communities associated to sterile plastic containers and kept at 4 °C
leaching piles was studied in order to select temperature before being processed [10].
tolerant microorganisms.
Chemical analysis of a representative sample
Fungi are one of the main components of of the leach pile material was carried out by
biota in soils and mineral substrates. Also, the instrumental analysis laboratory of the
under certain environmental conditions, Mining Research Institute (IIM), determined
they may be efficient biogeochemical by Inductively Coupled Plasma (ICP-OES
agents and metal bioaccumulators. Among Perkin Elmer 7300DV). In order to
them, Penicillium sp. is one of the most determine the mineral phases present in the
prominent genera in this methodology [8].

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sample, a steroscopic microscope Leica monitor growth development. Filamentous

S6D was used. fungi presenting different colony
morphotypes were isolated and mantained in
the same agar medium at 5 °C.

2.3. Identification of the isolations

For the molecular biology identification; the

selected fungi were individually grown in a
Sabouraud Dextrose medium at 30 °C for 72
h in an orbital shaker (250 rpm). The
mycelia were then collected by
centrifugation at 10000 × g (10 min),
suspended again in 2 M NaCl and finally
washed twice with sterile distilled water.
The washed mycelial mats were frozen in
liquid nitrogen and ground to powder by
using a sterile pestle and mortar. The DNA
from the ground mycelia were immediately
extracted with phenol: chloroform: - isoamyl
alcohol (25:24:1) and washed twice with
chloroform: - isoamyl alcohol (24: 1). Next,
two volumes of absolute ethanol and 0.1
volume of 3M potassium acetate were added
to the final aqueous phase in order to achieve
Figure 1. Hualilan Mine, Argentina, Satellite DNA precipitation, mixed by inversion and
photo.
then centrifuged (8000 ×g, 10 min, 4 °C).
The pellet was washed twice with 70 % (in
2.2. Isolation of filamentous fungi
weigt) ethanol, dried and finally suspended
again in sterile water. Sequencing of the
Heavy metal-tolerant filamentous fungi
genes nucleotides was performed by
were isolated using Sabouraud glucose agar
Macrogen (Korea). The sequences were
(Biopack) as culture medium, the
analyzed and edited when necessary by
composition of g/L is: animal digestion
using the Invitrogen Vector NTI Advance
pancreatic tissue (5), casein pancreatic
10.3.0 software (Invitrogen, San Diego, CA,
digestion (5); dextrose anhydrous (36.4)
USA). The identification of the strain was
and bacteriological agar (15).
carried out by comparison with the
A portion of 1 g of mining waste was
sequences of strain-type available in
weighed and sterile distilled water was
MycoBank by using the BLAST tool. The
added up to a final volume of 10 mL in a
sequences were also compared with the
test tube and then stirred until
species hypothesis in the UNITE 13 database
homogenization. Aliquots of 0.1 mL-
[11-13].
aliquots were withdrawn, serially diluted
and placed on plates on Sabouraud glucose
Macroscopic identification was performed
medium, in triplicate. Plates were incubated
according to a taxonomic scheme using
at 28 ºC for 24 h, and observed regularly to
MEA (in g/L: malt extract 30, peptone from

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soybean 3, bacteriological agar 15), CYA factorial arrangement 3x6x1, a Petri dish as
(in g/L: K2PO4 1, czapeck concentrate 10, an experimental unit and three repetitions.
yeast extract 5, sacarose 30, agar-agar 15 The growth diameters of each fungus at the
and traces of metal 1ml/L), AGS (in g/L: different concentrations (PRGI) were taken
animal digestion pancreatic tissue 5, casein as reference.
pancreatic digestion5; dextrose anhydrous
36.4 and bacteriological agar 15), PDA
(Potato Dextrose Agar, Biopack in g/L: 3. RESULT AND ANALYSIS
potato extract 4; glucose 20; bacteriological
agar 15), and G25N (K2HPO4 1 g/L, 3.1. Chemical analysis and visual
czapeck concentrate 10 ml/L) as culture identification of minerals of mining waste
mediums [14,15]. material

As a result of the chemical characterization

2.4. Heavy metal ions tolerance of the mine residues presented in Table 1,
iron is determined as the major component,
For this test, a PDA (Potato Dextrose Agar) whereas zinc, calcium, manganese,
commercial culture medium Biopack was aluminum, potassium and copper are
used, the composition of which is: potato secondary components. Regarding the
extract (4 g/L); Glucose (20 g/L); presence of heavy metals, lead is found in
bacteriologic agar (15 g/L). After being first place, followed by cadmium and silver,
sterilized in autoclave at 121 °C for 15 min, this last is present in a small proportion.
it was allowed to cool to a temperature of Moreover, the minerals from the pile showed
50 °C and, under sterile conditions, it was a leachate with a paste pH slightly basic
placed in Petri dishes, thereby obtaining 20 essentially due to the presence of carbonates
ml of culture medium waiting for gelation. in its composition. These mining wastes are
the result of the accumulation of mineralized
Various concentrations of each metal ion rocks which have been subjected to the
(25-800 ppm) were added. The metal salts process of alkalization with slacked lime,
(copper sulphate and lead nitrate) were used and later leached with sodium cyanide in
to prepare stock solutions (800 ppm). order to extract the valuable minerals.

Each plate was inoculated with a mycelial The visual identification of minerals of the
disk of the fungus by using a 0.5 cm mining waste suggested the presence of:
diameter punch previously sterilized. Plates pyrite (Py), Quartz (Qz), clays, mica, rock
without metals ions were also inoculated as clasts as well as of oxides (Figure 2), and
controls [16]. The plates were grown in a other supplementary material. In the sample,
culture oven at 28 °C. The mycelia growth a strong reaction to the addition of cold HCl
in each plate was observed during a whole was observed, which shows the presence of
period of the fungus development and then calcium carbonate.
compared with the control plate by
measuring in centimeters the growth For this reason, the chemical-mineralogical
diameter of each plate. All samples were characterization corresponds with that of
treated in triplicate. The statistical analysis minerals from a lixiviation pile, where the
was performed by using the Categorical protective alkalinity of a pH higher than 10
Multifactorial ANOVA, under a completely is achieved by adding measured quantities of
randomized experimental design with

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alkali; this is typical of a lixiviation process fungi capable of growing in those

with cyanide to dissolve gold and silver. contaminated environments were isolated.

Table 1. Data of elements determined through

ICP. 3.2. Identification of filamentous fungi
Cu Fe Cr Al Pb Si Na
(µg/g) (µg/g) (µg/g) (µg/g) (µg/g) (µg/g) (µg/g) As a result of this study, strains of the genera
1722 183400 7.90 10930 5045 172.3 3927 Fusarium and Penicillium were found (see
Table 2). According to Cárdenas, et al.,
Mn Ca Mg Zn P Cd Li
(µg/g) (µg/g) (µg/g) (µg/g) (µg/g) (µg/g) (µg/g) 2015, several genera of fungi were isolated
18370 108000 3418 35850 N/D 159.70 41.80
from rivers in México, namely Alternaria sp,
Penicillium sp, Curvularia sp, Aspergillus
K Mo Ag Sr Ti Ni Co
(µg/g) (µg/g) (µg/g) (µg/g) (µg/g) (µg/g) (µg/g)
Niger, Aspergillus flavus, Cephalosporium
sp, Fusarium sp and Cladosporium sp [17].
5681 24.30 6.80 133.10 N/D 0.70 N/D
The isolated fungi show different patterns of
resistance to the metals analyzed: most grow
in lead and zinc, few in mercury and arsenic,
and the highest sensitivity found was that of
cadmium. These fungi can be used for the
uptake of heavy metals in solution either
alone or accompanied by other biomasses as
reported in the literature [17, 18]. A potential
heavy metal biosorbent of the fungal type of
the genus Penicillium sp. was isolated from
the contaminated soils of the El Alacrán
Mine (Colombia). The experimental results
showed that the fungus had a high capacity
to eliminate: Pb (II), Cd (II) and Hg (II) in
an aqueous solution [19]. Holanda et al.,
(2016), reported that a sample of soil from a
mine in the province of Chonnam in South
Korea which was contaminated with heavy
metals was an important source of
biosorbents. The chemical analysis showed
high contents of lead (Pb) at 357 mg/kg and
cyanide (CN) at 14.6 mg/kg in the soil. The
experimental results showed that Penicillium
Figure 2. (A-B). Image of mineralogy of leach sp. MRF – 1 was the best lead resistant
pile. fungus among the four species of fungus
tolerant to heavy metal isolated from the soil
It is considered that the chemical- [20]. Moreover, the literature reports the
mineralogical characterization of the isolation of a native strain identified as
mining waste is clear, as the final objective Klebsiella sp. 3S1 biofilm isolated from a
was to show the presence of heavy metals wastewater treatment plant in the old mining
in the mining waste, which was precisely district Linares-La Carolina located in Sierra
the place from where these filamentous Morena (España) [21].

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One strain of each species was macro and Access

Closet Match Identification
microscopically identified as detailed Fungi number
(UNITE) (%)
below: GenBank
SH182481.07F
Penicillium simplicissimum A: it has double U Penicillium
A EF634422 simplicissimum 100
whorl brushes and lanceolate phialides with (Samson et al.,
colonies of a diameter of less than 10 mm 2011)
after 7 days of incubation in G25N SH219102.07F
M EF453114 98.71
medium. Colonies in CYA medium (with a U Fusarium sp
diameter of 35-50 mm) are radially sulcate SH209380.07F
or have irregular flocculated wrinkles and U Penicillium
B2 GQ221866 funiculosum 100
dense, white-grey mycelium. Conidia (Samson et al.,
production is low to moderate. These are 2011)
greyish green to dull green, sometimes they
look more yellow or olive. The obverse of Penicillium funiculosum B2: colonies in
the colony was pale yellow. Colonies in CYA are approximately 250 mm in
MEA usually have a diameter of 35-45 mm, diameter; they are flat, crusted, hairy, and
and are floccose; they have a white or pale sulcate with dark red color; they produce
mycelium; the conidia are similar to those soluble reddish pigment and few transparent
of CYA in color; the reverse is pale, exudates. They show whitish to reddish
brownish, dark brown, dark green, or most conidia. Colonies in MEA are approximately
of the times pink, centrally or in sectors, 300 mm in diameter, creamy in appearance
and occasionally even bright red. Colonies and with reddish excrescences toward the
occur in G25N of 10-18 mm in diameter center. Although the front face of the colony
with typically flat, low, from soft to does not show sulci when seen from above,
floccose white or yellow mycelium, it presents well marked rugosities. It
greenish-grey conidia with a pale reverse. produces transparent to reddish exudates.
At 37 °C, colonies are 10-30 mm in Conidia are from whitish to greenish in
diameter, dense and hairy, with a clear color. Growth is almost undetectable at 37
exudate and brown soluble pigment, and °C; conidia do not grow in G25N or at 4 °C.
they have a pale yellow reverse. The They have complex double whorl
colonies show spherical conidia with conidiophores which are branched.
smooth walls, Figure 3 (a) and (b). Occasionally, they also have one-whorl
conidiomers; they have lanceolate phialides,
Fusarium sp M: genera characterized by the smooth stipes and rounded cones with
production of small hyaline, unicellular smooth walls, Figure 3 (e) and (f).
conidia, which are born individually from
solitary phyllodes. The conidia are born
sequentially, but they are not connected to 3.3. Tolerance to heavy metals
each other. They show variability in the
structures responsible for both, forming the The application of the analysis of variance
cells, and for asexual reproduction. The (ANOVA) is shown hereunder, where the
colony is white in ASG medium, but pink variability associated with each factor, the
in PDA medium, Figure 3 (c) and (d). interaction among them, and the statistically
significant terms are studied. In Table 3, the
Table 2. Molecular identification of fungi
isolated from a leach pile.

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case of lead is shown, and in Table 4, the

case of copper is shown.

Figure 3. Macroscopic morphology of Penicillium simplicissimum colony in G25N medium (a). Detail of
spherical conidia in optical microscope with phase contrast at 40× magnification (b). Macroscopic
morphology of Fusarium sp. colony in ASG medium (c); Detail of conidia in optical microscope with
phase contrast at 40 × magnification (d). Macroscopic morphology of Penicillium funiculosum colony in
MEA medium (e). detail of conidiophore in optical microscope with phase contrast at 40 × magnification
(f).

Table 3. Analysis of Variance to PRGI for the metal lead – Sum of Squares Type III
Source Sum of squares d.f Mean Square F- Value P-Value
MAIN EFFECTS
A:Fungus 10728.4 2 5364.18 37.81 0.0000
B:concentration of Pb 3717.4 6 619.566 4.37 0.0144
RESIDUALS 1702.39 12 141.866
TOTAL (CORRECTED) 16148.1 20

Table 4. Analysis of Variance to PRGI for the metal copper – Sum of Squares Type III

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Source Sum of squares d.f Mean Square F- Value P-Value

MAIN EFFECTS
A:Fungus 13750.3 2 6875.17 23.51 0.0001
B:concentration of Cu 10295.0 6 1715.83 5.87 0.0046
RESIDUALS 3508.8 12 292.4
TOTAL (CORRECTED) 27554.1 20

The ANOVA tables decompose the Table 5. Tolerance of Fusarium sp M,

variability of PRGI in contributions due to Penicillium funiculosus B2 and Penicillium
various factors. Taking into account the fact simplicissimum A to heavy metals and
that the Sum of Squares Type III was assessment of tolerance capacity expressed in
chosen, the contribution of each factor is PRGI.
measured by eliminating the effects of the M-H1 B2-H2 A-H3
Metal
other factors. The values-P proves the concentration
Fusarium Penicillium Penicillium
sp funiculosum simplicissimum
statistical significance of each of the factors (ppm)
Values of PRGI
for the metals studied, as they were lower Pb 25 15.60 49.87 0.00
than 0.05. Therefore, it can be concluded Pb 50 16.67 50.67 0.00
that these factors have a significant statistical Pb 75 19.02 53.33 0.00
effect over PRGI with a confidence level of Pb 100 20.00 54.67 0.00
95.0%. Pb 400 32.25 54.67 6.40
Pb 600 53.33 70.00 7.52
As can be seen in Table 5, the strains Pb 800 85.33 79.60 11.63
showed adequate tolerance to Cu (II) and Pb Cu 25 0.00 36.00 0.00
(II) at various concentrations. The Cu 50 0.00 59.20 0.00
Percentage of Radial Growth Inhibition Cu 75 17.87 63.60 0.00
(PRGI) is also shown in this table. For this Cu 100 22.27 78.40 0.00
purpose, a range of growth has been taken as Cu 400 35.56 83.56 5.23
reference, starting with the initial diameter Cu 600 78.27 91.60 8.02
of the inoculum of each fungus until Cu 800 86.67 100.00 10.50
reaching its total development, which, at
most, is the extent of the complete plate; i.e. In particular, the PRGI values of the present
the initial proportion of inoculum was 0.5 study showed that the Penicillium
cm and the final ratio growth extent of the simplicissimum strain was the most tolerant
fungus in the whole of the plate was 4.25 to both metals, exhibiting zero radial
cm. inhibition percentages for the lowest
concentrations (from 25 ppm to 100 ppm) of
A value of PRGI = 0 % corresponds to the both lead and copper. Statistically,
total tolerance of the fungus to the metal, significant tolerance differences (PRGI)
while a PRGI value = 100 % corresponds to exist (P=0.00) in all the strains from which
fungi not tolerant to metal. the Penicillium simplicissimum A was the
fungus showing the highest tolerance (lowest
It may be claimed that out of the two metals PRGI), as shown in Figures 4 and 5,
assayed at various concentrations, lead was compared with the other two fungi. For the
the most highly tolerated by the three strains, case of lead at a concentration of 800 ppm,
even more than copper (Pb > Cu). the PRGI value is 11.63% whereas for Cu at
the same concentration, is 10.50%. The
method currently used to discriminate

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between the means is the Duncan’s multiple strategies for habitats contaminated with
comparison procedure. The results obtained heavy metals through various detoxification
in this study with this method were that the mechanisms such as biosorption the purpose
Pb concentrations with the lowest and also of which is to be used for the design of
homogeneous PRGI, did not give significant bioremediation processes. These offer the
differences in the PRGI values, and were the advantage of having cell wall components
lowest ones in the 25 to 400 ppm range. that exhibit excellent metal binding
Those producing significant differences were properties [24-26].
those with the highest metallic
concentrations, that is, from 600 to 800 ppm.
Moreover, the Cu concentrations with the
lowest PRGI and more homogeneous, which
did not have significant differences in the
PRGI values, were those within the 25 to
100 ppm range, whereas those producing
differences, i.e., they had significantly high
PRGI values, were those from 600 to 800
ppm. From these results it was possible to
establish the following: both for copper and Figure 4. PRGI graph versus the three fungi
lead, the highest metallic concentrations, isolated for lead concentrations: M-H1, B2-H2,
which is from 600 to 800 ppm, have and A-H3.
significantly high values of PRGI. The
statistical residue analysis showed that the
wastes had a normal behavior and were
independent.

The Penicillium funiculosum fungus shows a

better performance in smaller concentrations
of lead and copper up to 50 ppm with respect
to higher concentrations of both metals
respectively, where it shows a limited
performance. Moreover, Penicillium Figure 5. PRGI graph versus the three fungi
funiculosum is more tolerant to higher isolated for copper concentrations: M-H1, B2-
concentrations of lead than of copper, and is H2, and A-H3
not tolerant to the highest concentration of
this latter metal (800 ppm). In conclusion, taking into account the above
data, it can be said that for a PRGI equal to
The Fusarium sp fungus is tolerant to zero or near zero it can be stated that the
concentrations of up to 600 ppm of lead and fungus is tolerant or highly tolerant to heavy
400 ppm of copper while it is not so at the metals, in this case lead and copper, which
highest concentrations (600 and 800 ppm) of means that it can be used in the process of
both metals. This is consistent with the fact bioremediation. The type of technique used
that fungi are a versatile biosorption group in this study allows identifying and selecting
as they can grow under extreme conditions which is the most tolerant fungus.
of pH, temperature and nutrient availability
and high metal concentration [22,23]. 4. CONCLUSION
Microorganisms have developed survival

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 N. Hidalgo, P. Fernández, D. Bustos, M. Rosa, A. Senese
Revista Colombiana de Materiales, ISSN 2256-1013, No 18, pp. 21-32, 2021, https://doi.org/10.17533/udea.rcm.n18a02

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