Open Chemistry 2022; 20: 1096–1109
Research Article
Narda Fajardo Vidal*, Jorge Wong Dávila
Lead and cadmium removal with native yeast
from coastal wetlands
https://doi.org/10.1515/chem-2022-0211
received July 9, 2022; accepted September 4, 2022
Abstract: Water bodies affected by heavy metals have
been characterized in some natural ecosystems such as
coastal wetlands in Peru. For this reason, in the present
study, the determination of heavy metals lead (Pb), cad-
mium (Cd), and others was carried out in the water bodies
of the Regional Conservation Area (RCA) Wetlands of
Ventanilla using the Induction Coupled Plasma method.
Water samples were collected at the six most critical
stations for Pb and Cd, for the isolation of lead-tolerant
microorganisms in 2022 with the aim of evaluating native
microorganisms with removal potential of Pb and Cd. Yeasts
such as Candida guilliermondii, Candida famata, Cryptococcus
laurentii, Cryptococcus humicola, and Rhodotorula mucilagi-
nosa with tolerance to high concentrations of Pb were iso-
lated. The yeast with the best Pb tolerance result was Candida
guilliermondii isolated from groundwater (piezometer sam-
pling J1); Pb sorption was conducted with active yeast
(living biomass), whereas both Pb and Cd sorption were
conducted with inactive yeast (dead biomass). The results
were compared with those of a reference standard yeast
strain Saccharomyces cerevisiae: the native yeast proved to
have optimum behavior for the process.
Keywords: heavy metals, biosorption, yeast, wetland
1 Introduction
Wetlands are highly productive ecosystems that provide
a number of services of significant value to mankind. Flood
control, groundwater replenishment, sediment retention,

* Corresponding author: Narda Fajardo Vidal, Department of
Bioprocesses, Faculty of Chemistry and Chemical Engineering,
National University of San Marcos UNMSM, Calle Germán Amézaga
375, 15081 Lima, Peru, e-mail: narda.fajardo@unmsm.edu.pe
Jorge Wong Dávila: Department of Chemical Engineering, Faculty of
Chemical and Textile Engineering, National University of
Engineering UNI, Av. Tupac Amaru 210, 15333 Lima, Peru
Open Access. © 2022 Narda Fajardo Vidal and Jorge Wong Dávila, published by De Gruyter.
Attribution 4.0 International License.
water purification, recreation, as well as climate change
mitigation and adaptation, are just a few of the many valu-
able ecosystem services that wetlands provide [1].
Ecosystem services are natural assets produced by
the environment and utilized by humans such as clean
air, water, food, and materials and contribute to social
and cultural well-being [2].
In developing countries like those in Latin America,
the conservation of ecosystems is a critical issue. In Peru,
the Regional Conservation Area (RCA) Wetlands of Ventanilla
(located in the Callao Region) is one of many regional
habitat conservation systems protected by the Peruvian
Government through the Natural Resources and Environmental
Management Office.
One of the problems faced by the RCA Wetlands of
Ventanilla is the accumulation of heavy metals, particu-
larly lead (Pb), and cadmium (Cd), in its water bodies;
these metals can bioaccumulate in fish and affect birds
through the food chain. In addition to these metals, there
would be other sources of contamination in these wet-
lands, such as poorly disposed mining tailings deposits
in the buffer zone of the RCA Wetlands of Ventanilla.
On the other hand, it is known that among the
separation processes for the potential removal of heavy
metals from water, adsorption/biosorption could repre-
sent a technically and economically feasible mechanism
for the respective decontamination [3,4]. Moreover, sev-
eral studies have shown that the structural matrix of
particular biomass sources, including biomass from micro-
organisms, can translate into efficient systems for sorption
of heavy metals [5–10]. Particularly interesting is the appli-
cation of sustainable compounds from biomass such as
yeast (active or inactive), in whose organic and molecular
structure there are polysaccharides with sorbent capacity,
in addition to having a network of functional chemical
groups that can ultimately result in an efficient-capacity
global yeast for heavy metal sorption.
With the goal of proposing an alternative that repre-
sents a balance between biological [11] and chemical engi-
neering processes for the remediation of the wetlands in
question, a study was carried out to (a) determine the
presence of potential native yeast microorganisms in the
This work is licensed under the Creative Commons
 Lead and cadmium removal with native yeast from coastal wetlands
RCA Wetlands of Ventanilla and (b) evaluate the heavy
metal sorbent capacity of said native yeast structures.
When investigating the presence of heavy metals in the
aquatic environment, it was decided to isolate yeasts tol-
erant to high concentrations of lead from the most critical
sampling stations of the RCA Wetlands of Ventanilla eco-
system for bioadsorption tests in an experimental design
where yeast Candida guilliermondii isolated from the RCA
Wetlands of Ventanilla was compared with a commercial
strain of Saccharomyces cerevisiae. The objective was to
determine the biosorption efficiency of the native yeast
of the RCA Wetlands of Ventanilla to continue in a future
investigation with the removal of Pb from the ecosystem
surface water under laboratory conditions.
2 Materials and methods
2.1 Location of area of study
This research work was carried out at the RCA Wetlands of
Ventanilla. The protected natural area of the reserve is
approximately 275 ha.
An aerial view of the RCA Wetlands of Ventanilla;
then including all available sampling stations M-1, M-2,
M-3, M-4, M-5, M-6, M-7, M-8, M-9, M-10, M-11, M-12, M-
13, M-14, M-15, M-16, and M-17 is shown in Figure 1. Based
on results from former studies in the RCA Wetlands of
Ventanilla, six stations were chosen as ideal in order to
isolate microorganisms such as yeasts, actinomycetes
and environmental fungi. We focused our research on
yeasts that showed high tolerance to Pb (1, 5, 10 and
1,000 mg/L) in laboratory tests.
2.2 Sampling
In August 2019, water samples were taken at each of the
seventeen sampling stations established back in 2015,
in order to determine which heavy metals were the
most critical in the RCA Wetlands of Ventanilla eco-
system and to design a research project oriented to the
isolation of microorganisms with bioremedication poten-
tial. The values of Pb and Cd reported by Fajardo [12] in
the same 17 sampling stations were also considered. Of
the 17 sampling stations established back in 2015, 15 were
for surface water and 2 were for groundwater. Sampling
stations M-1, M-2, M-3, M-4, M-5, M-6, M-7, M-8, M-9, M-
10, M-11, M- 12, M-13, M-14, M-15 were surface water; of
 1097
these, sampling stations M-8 and M-9 correspond to sur-
face water channels. Two piezometers were also estab-
lished, which consisted of stations M-16 and M-17 (for
underground water). One simple sample was taken per sta-
tion for total metals by induction coupled plasma mass
spectrometry (ICP-MS) and 1 simple sample per station
was put in a 250 mL sterile glass bottle for microorganism
isolation procedure. Successful isolation was achieved in
2022, but (during the previous years samples were col-
lected to adjust and improve the yeast isolation protocol).
The sampling is representative of the RCA Wetlands of
Ventanilla ecosystem because the 17 stations evaluated
over the years are distributed in the permanent bodies of
water and in two groundwater facilities that were built in
2015 and that were kept under permanent maintenance so
as not to lose them. The criteria for establishing the sta-
tions considered the ecological importance as bird nesting
areas, proximity to the population, and the water mirrors
with access allowed only for research purposes (M-12). For
those stations that were implemented within the environ-
mental and safety safeguards required by Prociencia, a
mobile and assemblable wooden bridge was built, which
was only used during sampling.
For groundwater sampling, the work area around the
piezometer was prepared, and a plastic cover was placed on
the ground to prevent cross-contamination. As it was a mon-
itoring piezometer, the cover was removed and the piezo-
meter was purged. A suction system was used and a volume
of water equal to at least three (03) times the volume of the
piezometer was removed from the piezometer. During the
purging procedure, the parameter stabilization criterion was
applied, and only after they stabilized, sampling could
begin. A bailer was used for each piezometer. The bailer
was lowered gently until it came into contact with the sur-
face of the water, avoiding at all times that the sediments
were disturbed and that it came into contact with the walls
of the piezometer, then the sample was collected. Once pre-
served, hermetically covered, and labeled, the samples were
placed in coolers, in vertical position, with their respective
cooling gels, at an approximate temperature of 4°C, and
with their corresponding chain of custody.
The sampling of surface waters was carried out in
non-turbulent zones, with a uniform current. Sampling
at the surface and bottom of the surface water body was
avoided. The bottles were submerged below the surface
of the water to a depth of 20–30 cm, after which they were
turned so that the mouth pointed toward the current (if it
existed), or by creating said current by dragging the
bottle on the inside of the water. For the sampling of total
metals, the container was rinsed with the water to be
sampled, at least twice before taking the sample.
1098  Narda Fajardo Vidal and Jorge Wong Dávila

Figure 1: Location of available sampling stations M-1, M-2, M-3, M-4, M-5, M-6, M-7, M-8, M-9, M-10, M-11, M-12, M-13, M-14, M-15, M-16,
and M-17 at the Ventanilla Wetlands Regional Conservation Area (RCA Wetlands of Ventanilla), Callao, Peru, established in 2015. Sampling
stations J1, J2, J3, J4, J5, and J6 were sampled in this work.
 Lead and cadmium removal with native yeast from coastal wetlands  1099

The six stations selected for the 2022 sampling were
chosen considering the historical data of Pb and Cd
values reported in February, May, and August 2015 and
August 2019 (Figures 2 and 3).
Water samples were collected at seventeen sampling
stations in order to carry out the chemical characteriza-
tion of Pb. The 17 sampling stations were those estab-
lished in 2015. Of the six sampling stations chosen, five
were for surface water and one was for groundwater (J1);
the groundwater samples were taken from piezometers.
The six sampling stations selected were: M-1, M-4, M-5,
M-10, M-15, and M-17 that were named for microbiolo-
gical studies such as J1, J2, J3, J4, J5, and J6; the collected
samples were seeded in 250 mL flasks with 100 mL YPG
broth [13] for the isolation of microorganisms tolerant to
heavy metals.
the General Analytical Services Environmental Laboratory
SAG SAC (Lima, sagperu.com), accredited by the INDEC-
OPI – National Institute for the Defense of Competition and
Protection of Intellectual Property, where the samples
were analyzed using ICP-MS, and in the six stations of
sampling selected for the isolates of microorganisms (J1,
J2, J3, J4, J5, and J6), samples were taken in 250-mL sterile
glass bottles for the microbiological isolates in the labora-
tory and samples in plastic bottles for total metals, pH,
temperature, conductivity, and dissolved oxygen were
also measured in the field in order to better characterize
the water from the sources of isolation of microorganisms.
2.4 Isolation of microorganisms

Isolation was performed in YPG broth of 5 g/L yeast

2.3 Harvest
The amount of each sample was 250 mL water, in new
plastic containers, properly labeled. A HNO3 solution was
added until pH <2 for the preservation of total metals; the
samples were immediately frozen and were then sent to
extract, 10 g/L meat peptone, 20 g/L glucose, and 18 g/L
agar (HiMedia Laboratories, Maharashtra/Sigma-Aldrich,
St. Louis MO) with 0.02% chloramphenicol (Portugal
brand of Peruvian origin, labportugal@laboratoriospor-
tugal.com) at pH 5.5. The ingredients to formulate the
yeast culture media were from Himedia, and the rest of
the reagents used in the study were obtained from Sigma-

Pb in RCA Wetlands of Ventanilla

0.10
0.09
0.08
0.07
0.06
mg/l

0.05
0.04
0.03
0.02
0.01
ECA = 0,0025
0.00
M-1
M-2
M-3
M-4
M-5
M-6
M-7
M-8

Pb_Ago-2019
M-9
M-10
M-11
M-12
M-13
M-14

Pb_Feb-2015
M-15

M-16

M-17

Locaon
Pb_Feb-2015 Pb_May-2015 Pb_Ago-2015 Pb_Ago-2019 Pb_Ene-2022

Figure 2: Lead values reported in 2015 [12], the values obtained in 2019 and the results of the 6 sampling stations J1, J2, J3, J4, J5, and J6
of 2022.
1100  Narda Fajardo Vidal and Jorge Wong Dávila

Cd in RCA Wetlands of Ventanilla

0.009
0.008
0.007
0.006
mg/l

0.005
0.004
0.003
0.002
0.001
ECA = 0,00025
0.000
M-1
M-2
M-3
M-4
M-5
M-6
M-7
M-8
Cd_Ago-2019

M-9
M-10
M-11
M-12
M-13
M-14
Cd_Feb-2015

M-15
M-16
M-17
Locaon

Cd_Feb-2015 Cd_May-2015 Cd_Ago-2015 Cd_Ago-2019 Cd_Ene-2022

Figure 3: Cadmium values reported in 2015 [12], the values obtained in 2019 and the results of the 6 sampling stations J1, J2, J3, J4, J5, and J6
of 2022.

Aldrich, St. Louis MO; the isolation system was in flasks
of 250 mL capacity, with 100 mL of YPG broth at 150 rpm
of agitation and at 24 degrees centigrade, two flasks per
sampling station were prepared, six flasks named (J1, J2,
J3, J4, J5, and J6) and the six flasks with YPG broth sup-
plemented with 0.4 mg/L of lead from Sigma Aldrich
brand lead nitrate salt, these systems were named (J1-
JJJ, J2-JJJ, J3-JJ, J4-JJJ, J5-JJJ, and J6-JJJ). Of the 12 flasks
of the protocol, 0.1 mL of process broth was seeded by
dissemination with a drigalski spatula in Petri dishes
with YPG agar. The plating procedure was performed at
24, 48, 72, 96, and 120 h of the process; the flasks with the
samples were kept under constant agitation. Suspicious
and interesting colonies were isolated in YPG broth at pH
5.5 to later generate pure cultures, and gram stains were
also performed. Among the isolated microorganisms, we
obtained some actinomycetes, environmental fungi, and
nine native yeasts.
analysis kit API® ID 32C (Biomèrieux, Marcy-l'Étoile),
which is a standardized system for the identification of
yeasts composed of 32 assays of miniaturized assimila-
tion. The identification output was coupled to a corre-
sponding computer program, and the identified yeasts
were Candida guilliermondii, Candida famata, Cryptococcus
laurentii, Cryptococcus humicola and Rhodotorula mucilagi-
nosa (Table 3); the yeasts were seeded in tubes with YPG
broth at concentrations of 1, 5, 10 and 1,000 mg/L of lead
and incubated at 24°C for 1 week. Readings were taken every
day in order to determine if there was growth or inhibition
(Table 4). The identified yeasts were transferred to cepario
in cryovials with YPG broth supplemented with 10% vol
glycerol and stored at −20°C.
2.6 Total metal analysis

Metal concentration was analyzed using ICP-MS. For the

2.5 Identification of microorganisms
Nine wetland yeasts with typical morphology observed
in the gram staining of the smears of the pure colonies
were identified and biochemically identified by means of
ICP-MS analyses of the samples from the flasks of the
experimental design, the most efficient active yeast in Pb
removal was Candida guilliermondii (isolated from the wet-
land piezometer) and was compared with Saccharomyces
cerevisiae (pattern commercial strain); 10 mL aliquots were
taken in each sampling at six different time intervals, put in
 Lead and cadmium removal with native yeast from coastal wetlands  1101

Table 1: Concentration of Pb and Cd in bodies of water in the Ventanilla Wetlands Regional Conservation Area (RCA Wetlands of Ventanilla)
during 2022

Samples Sampling station Pb (mg/L) January 2022 Pb-ECA 2017 Cd (mg/L) January 2022 Cd-ECA (mg/L) 2017

J1 Piezómetro 0.0257 0.0025 0.00103 0.00025

J2 Espejo Rojo 0.0066 0.0025 0.00021 0.00025
J3 Cañaveral 0.0040 0.0025 0.00019 0.00025
J4 Canal Cerco 0.0123 0.0025 0.00015 0.00025
J5 Filtro de agua 0.0110 0.0025 0.00076 0.00025
J6 Pisciplaya 0.0002 0.0025 0.00009 0.00025

Concentration expressed in mg/L.

15 mL falcon tubes, centrifuged, and the supernatants were
analyzed on Pb content to evaluate the percentage of lead
removal in six time intervals. The flasks started with 1 mg
Pb/L in YPG broth at pH 5 and 6, the process times were at 3,
7, 11, 24, 48 and 72 h, and the reaction took place in 250 mL
flasks filled with 100 mL suspension each, at 150 rpm agitation
speed and 30°C temperature. The set points were controlled
by a shaker incubator, model TOU-120 (MRC Laboratory-
Instruments, Holon, www.mrclab.com). The results of removal
of the active yeasts are shown in Figure 4 for the native yeast
Candida guilliermondii and Figure 5 for the commercial baker’s
yeast Saccharomyces cerevisiae.
For the biosorption by inactivated yeast biomass, 1 g
of inactivated yeast was applied at different thermal
treatment procedures: at 45 and 121°C, drying after cen-
trifugation of the biomass generated for 48 h in YPG broth
and their respective washing with sterile distilled water
followed by second centrifugation. The drying was carried
out in a forced-air circulation oven, model UF110 (Memmert
GmbH + Co. KG, Schwabach, www.memmert.com) at 45°C
for 24 h for all treatments; of the biomass generated in YPG
broth for 48 h growth for both yeasts, in the heat treatment
at 121°C for 30 min in an autoclave, the biomass was cen-
trifuged in 50 mL falcon tubes (Brand ISOLAB GmbHha of
German origin, isolabgmbh.com), at 2,600 rpm for 10 min;
the supernatant was discarded, and the sediment was sus-
pended in sterile distilled water to proceed to second
centrifugation as washing. The supernatant was again dis-
carded, and the washed sediment was dried at 45°C in the
oven for 24 h.
For the Pb and Cd biosorption system, 1 g of inacti-
vated yeast was used in 250 mL flasks filled with 100 mL
deionized water at pH 6 with an initial lead concentration
of 3.2814 and 0.17135 mg/L of cadmium; 10 mL samples
were taken in 15 mL falcon tubes at 1, 3, 24 and 48 h.
The samples were centrifuged, and the supernatant was
analyzed using ICP-MS. In other investigations in which
Saccharomyces cerevisiae AUMC 3875 was evaluated for
live and dead biomass, the maximum capacities for lead(II)
absorption were reached at pH 5.0, initial concentration of
metal ions 300 mg/L and biomass dose 3 g/L. Maximum
biosorption was reached after 3 h and 20 min for live and
dead cells, respectively [14].
For biosorption as inactivated yeasts in water from
the Ventanilla Wetlands Regional Conservation Area, 1
gram of inactivated yeast was worked at 121°C, drying
after centrifugation of the biomass generated for 48 h in
YPG broth and their respective washing with sterile dis-
tilled water and second centrifugation was carried out in
a Memmert model UF110 oven at 45°C for 24 h for all
treatments; of the biomass generated in YPG broth for
48 h of growth for both yeasts, in the heat treatment at
121°C for 30 min in an autoclave, the biomasses were
centrifuged in 50 mL falcon tubes at 2,600 rpm for

Table 2: Physicochemical parameters

Samples Sampling station pH Temperature (°C) Dissolved oxygen Conductivity (mS/cm) Salinity (%)

J1 Piezómetro 8.87 22 4.78 37.6 23.8

J2 Espejo Rojo 9.83 33.1 15.76 86.5 >42
J3 Cañaveral 9.52 33.3 12.73 21.9 12.82
J4 Canal Cerco 9.09 28.1 10.3 14.8 8.59
J5 Filtro de agua 8.72 27.6 7.31 15.4 8.8
J6 Pisciplaya 8.64 28.2 9.08 18.49 10.94

pH, temperature, dissolved oxygen, conductivity, and salinity in bodies of water in the Ventanilla Wetlands Regional Conservation Area
during 2022. Concentration expressed in unit. pH, °C, mg/L, mS/cm year %, respectively.
1102  Narda Fajardo Vidal and Jorge Wong Dávila

1.2

1.0

0.8

0.6
[Pb] / mg/L

0.4

0.2

0.0
0 10 20 30 40 50 60 70 80
Time / h

pH 5 pH 6

Figure 4: Pb concentration during sorption by active biomass Candida guilliermondii.

1.2

1.0

0.8
[Pb] / mg/L

0.6

0.4

0.2

0.0
0 10 20 30 40 50 60 70 80
Time / h

pH 5 pH 6

Figure 5: Pb concentration during sorption by active biomass Saccharomyces cerevisiae.

10 min; the supernatants were eliminated, and the sedi-
ments were suspended in sterile distilled water to proceed
to second centrifugation as washing. The supernatant
was then eliminated, and the already washed sediments
were dried at 45°C in an oven for 24 h. For the lead and
cadmium biosorption system in water from the RCA Wetlands
of Ventanilla, 1 gram of inactivated yeast was used in 250 mL
flasks with 100 mL of water from the RCA Wetlands of
Ventanilla, at 30°C at 150 rpm system in a TOU-50/120
series incubator shaker (from Israel) and with pH 6; the
 Lead and cadmium removal with native yeast from coastal wetlands  1103

granulometry of the inactivated yeasts was D50 of 0.500 lead (mg/L) and cadmium (mg/L) ions in the solution,
millimeters, for which the No. 35 U.S. Mesh sieve was used, before and after biosorption, respectively; M is the mass
the removal system was with a concentration of lead initial of the biosorbent (g) lead and cadmium (mg/L), and V is
0.0134 mg/L and for cadmium 0.00033 mg/L; 10 mL sam- the volume of the solution (L).
ples were taken in 15 mL falcon tubes at times of 1, 2, 3, 48
(C0 − Ce) × V
and 72 h. The samples were centrifuged, and the super- qe = , (1)
M
natant was analyzed using ICP-MS.
(C0 − Ce)
%R = × 100. (2)
C0

2.7 Adsorption isotherms
To obtain the adsorption isotherm, a procedure was car-
ried out maintaining a fixed amount of bioadsorbent of
1 g in 100 mL and varying the initial concentrations of
lead and cadmium in the range of 0.1–1 mg/L of Pb and
0.01–0.1 mg/L of Cd. In each flask, the mixture of the lead
and cadmium solution with the biosorbent was kept
under stirring for the equilibrium time determined in
4 h. All other parameters were kept constant (pH 6 and
150 rpm). With the results of the concentration of lead
and cadmium adsorbed during biosorption, the values of
the biosorption capacity at equilibrium (qe) were plotted
against the final concentration of lead at equilibrium (Ce);
in this way, the isotherm was obtained biosorption at equi-
librium. In all cases, at the end of the contact time of lead
and cadmium with the bioadsorbent, the solid and liquid
phases were separated by filtration, using filter paper.
The liquid phases containing the residual concentration
of Pb and Cd were analyzed using ICP-MS. The amount of
lead ions retained by the biosorbent or biosorption capa-
city (qe, mg/g) and the removal percentage (R%) were
calculated using equations (1) and (2), respectively, where
Co and Ce are the initial and equilibrium concentration of
The Freundlich isotherm model was the one that
best fit the results obtained in the experiment for both
yeasts.
1
log Qe = log Kf + log Ce.
n
2.8 Data processing
To evaluate the results of each of the sampling stations,
tables were generated with the values obtained in each
environmental monitoring, and the values of lead and
cadmium in water are declared in Table 1.
The values of pH, temperature, dissolved oxygen,
conductivity, and salinity of the six sampling stations
are detailed in Table 2.
The yeasts identified by Kit API® ID 32 C are shown in
Table 3, where the sampling station from which they were
isolated and the culture broth used are specified.
The identified yeasts and their tolerance to different
Pb concentration values are shown in Table 4, where the
sign +, ++, or +++ was placed according to the degree of
turbidity observed in tubes with YPG broth with different
concentrations.

Table 3: Identification of yeasts isolated from the RCA Wetlands of Ventanilla in 250 mL flasks with 100 mL of YPG broth and in YPG broth
with lead

Sampling station Broth Strain Gram stain Identification by API® 32C

Cepa commercial YPG Saccharomyces cerevisiae Yeast Saccharomyces cerevisiae

J1 (M-17) YPG J1 Yeast Candida guilliermondii
J1 (M-17) YPG + Pb J1-JJJ Yeast Candida guilliermondii
J2 (M-10) YPG Not isolated — —
J2 (M-10) YPG + Pb Not isolated — —
J3 (M-4) YPG J3 Yeast Candida famata
J3 (M-4) YPG + Pb J3-JJJ Yeast Criptococcus laurentii
J4 (M-5) YPG J4 Yeast Rhodotorula mucilaginosa
J4 (M-5) YPG + Pb J4-JJJ Yeast Criptococcus laurentii
J5 (M-1) YPG J5 Yeast Candida famata
J5 (M-1) YPG + Pb J5-JJJ Yeast Rhodotorula mucilaginosa
J6 (M-15) YPG J6 Yeast Criptococcus humicola
J6 (M-15) YPG + Pb Not isolated — —
1104  Narda Fajardo Vidal and Jorge Wong Dávila

Table 4: Yeasts isolated from RCA Wetlands of Ventanilla and degree of tolerance to different concentrations of Pb

Pb concentration in YPG broth (mg/L) 1 5 10 1,000

Code Strain Temperature (°C) pH 5 pH 6 pH 5 pH 6 pH 5 pH 6 pH 6

CC Saccharomyces cerevisiae 24 +++ +++ +++ +++ +++ +++ +++

J1 Candida guilliermondii 24 +++ +++ +++ +++ +++ +++ +++
J1-JJJ Candida guilliermondii 24 +++ +++ +++ +++ +++ +++ +++
J3 Candida famata 24 +++ +++ +++ +++ +++ +++ ++
J3-JJJ Criptococcus laurentii 24 ++ ++ + + + + +
J4 Rhodotorula mucilaginosa 24 +++ +++ +++ +++ +++ +++ ++
J4-JJJ Criptococcus laurentii 24 + + + + + + +
J5 Candida famata 24 +++ +++ +++ +++ +++ +++ ++
J5-JJJ Rhodotorula mucilaginosa 24 +++ +++ +++ +++ +++ +++ ++
J6 Criptococcus humicola 24 +++ +++ +++ +++ +++ +++ +++

3 Results and discussion
The analysis of the water samples showed values above
the maximum permissible limit for the conservation of
aquatic environments in some of the sampling stations
(see Table 1). The lead and cadmium values of the RCA
Wetlands of Ventanilla exceeded or were close to the
national environmental quality standards for water in
Peru, according to the standard established by the Min-
istry of the Environment; then, according to Peruvian
regulations [15] for water intended for conservation of
the aquatic environment (lakes – lagoons), the values found
were above the standards, the limits are for lead (standard
0.0025 mg/L), cadmium (standard 0.00025 mg/L), and
cadmium did not exceed the maximum permissible limits
in most of the stations evaluated, just in two stations: J1
(Piezómetro) y J5 (Filtro de agua).
The most critical heavy metal in the RCA Wetlands of
Ventanilla ecosystem was lead (0.0257 mg/L) at station J1, the
problem of cadmium (0.00103 mg/L) at station J1, (0.00076
mg/L) in the stationJ5; piezometer (J1) also showed high
values of lead and was the sampling station from which
Candida guilliermondii (strain J1-JJJ) was isolated.
From the isolation of yeasts tolerant to high concen-
trations of lead and their identification, Candida guillier-
mondii was selected for presenting the best results of

3.5 0.18

0.16
3.0
0.14
2.5
0.12
[Cd] / mg/L
[Pb] / mg/L

2.0 0.10

1.5 0.08

0.06
1.0
0.04
0.5
0.02

0.0 0.00
0 10 20 30 40 50 60
Time / h
Pb (45) Pb (121) Cd (45) Cd (121)

Figure 6: Pb and Cd concentrations during sorption by inactive biomass Candida guilliermondii at pH 6 and for two different thermal
biomass deactivation treatments: 45 and 121°C.
 Lead and cadmium removal with native yeast from coastal wetlands
3.5
3.0
2.5
[Pb] / mg/L
2.0
1.5
1.0
0.5
0.0
0 10 20
Pb (45)
Figure 7: Pb and Cd concentrations during sorption by inactive biomass Saccharomyces cerevisiae at pH 6 and for two different thermal
biomass deactivation treatments: 45 and 121°C.
tolerance to high concentrations of Pb, growth at dif-
ferent pH, and its reports of biotechnological applications
[16], due to the particular source of isolation at the RCA
Wetlands of Ventanilla, namely from groundwater. The
strain was compared with a commercial Saccharomyces
cerevisiae yeast, which, according to the state-of-the-art,
is one of the most effective strains in removing Pb. Both
yeasts were evaluated in an experimental design matrix as
active yeasts, the initial Pb concentration was 1 mg/L in
the flask system with YPG broth. Similar Pb sorption trends
were obtained for both yeasts in the same time intervals.
The Pb and Cd sorption trends were also studied for
dead biomass inactivated at 45 and 121°C. The initial Pb
and Cd concentration was 3.2814 and 0.17135 mg/L respec-
tively, in flasks with deionized water at pH 6, loaded each
flask with 1 g inactivated yeast.
In former studies, the highest Pb and Cd uptake was
obtained with Saccharomyces cerevisiae. The Pb biosorp-
tion capacity of all kinds of biomass was higher than that
of Cd [17]. For inactivation, the biomass of both yeasts
was generated separately in YPG broth at pH 6. At 48 h
growth in YPG broth, centrifugation was carried out in
50 mL falcon tubes at 3,600 rpm for 5 min; the superna-
tants were removed and the sediments were suspended. in
sterile distilled water to proceed to second centrifugation
 1105
0.20
0.18
0.16
0.14
0.12
[Cd] / mg/L)
0.10
0.08
0.06
0.04
0.02
0.00
30 40 50 60
Time / h
Pb (121) Cd (45) Cd (121)
as washing. The supernatant was then removed, and the
already washed sediments were dried at 45°C in an oven
for 24 h. The inactivated yeasts were ground and sieved
with US MESH number 35 of 0.5 mm, generating granulo-
metry D50 (50% passing and 50% retained), the lead and
cadmium removal trend graphs were similar for the first
three times of 1, 3, and 24 h in both inactivated yeasts. In
our research, we evaluated biosorption of lead and cad-
mium in an aqueous solution of the yeast Saccharomyces
cerevisiae and Candida guilliermondii inactivated at 45
and 121°C; obtaining the best results in the first 24 h of
contact where the lead values decreased to 0.3412 and
0.1159 mg/L and the cadmium values to 0.1159 mg/L at
24 h and 0.0106 mg/L at the time of contact with Candida
guilliermondii at 45 and 121°C, respectively (Figure 6).
For Saccharomyces cerevisiae, the best results were
also at 24 h of contact where lead values decreased to 0.0318
and 0.0817 mg/L and cadmium values to 0.01519 mg/L at
24 h and 0.01353 mg/L at 24 h. 3 h of contact at 45 and
121°C, respectively (Figure 7); in other investigations, good
inactivation results were reported at 45°C for Saccharomyces
cerevisiae [18].
Thermal treatment at different temperature values (to
get the inactivated yeasts = dead biomass) had a signifi-
cant effect on Pb removal using Candida guilliermondii
1106  Narda Fajardo Vidal and Jorge Wong Dávila

0.016

0.014

0.012

0.01
[Pb] / mg/L

0.008

0.006

0.004

0.002

0
control 1h 2h 3h 48h 72h
Time / h

Saccharomyces cerevisiae Candida guilliermondii

Figure 8: Pb Concentration during sorption by inactive biomass Saccharomyces cerevisiae and Candida guilliermondii from RCA Wetlands of
Ventanilla water sample.

and Saccharomyces cerevisiae similar to those reported
on Cr removal by Saccharomyces cerevisiae [19].
Other studies [20] report Mediterranean forested wet-
lands as good sites for yeast isolation for dye bioremediation
and highlight the importance of conserving these ecosystems.
Among yeasts, the most studied are Candida, Pichia, Crypto-
coccus, and Saccharomyces [21,22]. This research reports
Candida guilliermondii as a native yeast of the RCA Wet-
lands of Ventanilla with biosorbent properties for Pb and Cd.
It could be seen that the optimum pH for Pb adsorption
by active Candida guilliermondii and active Saccharomyces
cerevisiae was 5 and 6, respectively. In previous research,
pH 5 was reported as optimum for Saccharomyces cerevisiae
in removal processes [23].

0.00035

0.00030

0.00025
[Cd] / mg/L

0.00020

0.00015

0.00010

0.00005

0.00000
control 1h 2h 3h 48h 72h
Time / h
Saccharomyces cerevisiae Candida guilliermondii

Figure 9: Cd concentration during sorption by inactive biomass Saccharomyces cerevisiae and Candida guilliermondii from RCA Wetlands of
Ventanilla water sample.
 Lead and cadmium removal with native yeast from coastal wetlands  1107

0.00
-3.50 -3.00 -2.50 -2.00 -1.50 -1.00 -0.50 0.00
-0.50
-1.00
logQeq / - -1.50
y = 1.1326x + 0.2931 -2.00
R² = 0.9913 y = 0.731x - 1.3008
R² = 0.9913 -2.50
-3.00
-3.50
log[Cd]eq / -

J1-JJJ (D50 = 0.5 mm) / Freundlich fing

Saccharomyces sp. (D50 = 0.5 mm) / Freundlich fing
Linear (J1-JJJ (D50 = 0.5 mm) / Freundlich fing)
Linear (Saccharomyces sp. (D50 = 0.5 mm) / Freundlich fing)

Figure 10: Cadmium sorption model by inactive biomass Saccharomyces cerevisiae and Candida guilliermondii.

In the results of lead and cadmium removal (Figures 8
and 9) from water from the RCA Wetlands of Ventanilla
under laboratory conditions, using the yeasts Saccharo-
myces cerevisiae and Candida guilliermondii inactivated at
121°C in the 5 removal times of 1, 2, 3, 48, and 72 h, very close
results are obtained for both yeasts, considering that Sacchar-
omyces cerevisiae is a reference yeast in lead and cadmium
removal according to the study of the art; the results obtained
in Candida guilliermondii make it a good candidate as yeast
native inactivated for remediation work in the Ventanilla
Wetlands Regional Conservation Area.
Due to the removal results as active and inactivated
yeast, Candida guilliermondii is presented as a native
yeast with lead and cadmium removal potential for the
Ventanilla Wetlands Regional Conservation Area.
The sorption patterns of cadmium and lead by bio-
masses inactivated with D50 of 0.5 mm granulometry of
Saccharomyces cerevisiae and Candida guilliermondii respec-
tively are shown in Figures 10 and 11.
The Freundlich isotherm model was the one that
best fit the results obtained in the experiment for
Saccharomyces cerevisiae and Candida guilliermondii

0.00
-1.80 -1.60 -1.40 -1.20 -1.00 -0.80 -0.60 -0.40 -0.20 0.00
-0.50
logQeq / -

y = 0.9851x - 0.4425 y = 2.2561x + 0.3889 -1.00

R² = 0.9989 R² = 0.9993
-1.50

-2.00

-2.50
log[Pb]eq / -

J1-JJJ (D50 = 0.5 mm) / Freundlich fing

Saccharomyces sp. (D50 = 0.5 mm) / Freundlich fing
Linear (J1-JJJ (D50 = 0.5 mm) / Freundlich fing)
Linear (Saccharomyces sp. (D50 = 0.5 mm) / Freundlich fing)

Figure 11: Lead sorption model during sorption by inactive biomass Saccharomyces cerevisiae and Candida guilliermondii.
1108  Narda Fajardo Vidal and Jorge Wong Dávila
as yeasts inactivated at 121 degrees and dried at 100°C
in the 0.5 mm grain size.
No yeast was isolated from station J2, the pH was
9.83, and the salinity was extreme in the water sample,
being the impacted in lead (0.0066 mg/L) of all the sam-
pling stations.
4 Conclusions and
recommendations
This study has determined the presence of heavy metals
such as lead and cadmium in surface water bodies and
the one underground water stations J1 of the RCA Wetlands
of Ventanilla in the years 2022; the most critical heavy metal
in the ecosystem was lead with a maximum value of
0.0257 mg/L at station J1.
The sampling stations most affected by lead were
those established in the Piezómetro J1, en Espejo Rojo
J2, in the buffer zone “Cañaveral” J3, Canal Cerco J4
and Filtro de Agua J5; the most optimal yeast in lead
removal was Candida guilliermondii isolated from piezo-
meter J1 (groundwater).
Tolerance tests to different concentrations of lead
made it possible to evaluate the nine isolated native
yeasts in growth time, determining that some are slow-
growing, such as Cryptococcus laurentii (strain J3-JJJ);
however, Candida guilliermondii (J1-JJJ) presented toler-
ances to high concentrations of lead, optimal growth at
pH 5 and 6, a characteristic that allows the strain to be
maintained in different culture media and evaluated in
different experimental optimization designs.
The yeast Rhodotorula mucilaginosa (J5-JJJ) showed
high tolerance to different concentrations of lead, but its
growth was not optimal at pH 6 with a lead concentration
of 1 mg/L and it is a yeast that is frequently found in
natural ecosystems.
The yeast Candida guilliermondii was the one that
presented the best results of tolerance to different con-
centrations of lead, and its removal results are very
similar to those of Saccharloomyces cerevisiae (reference
yeast) evaluated as active yeasts in an experimental
design of Pb removal with three repetitions.
Applications in removal of lead and cadmium by the
yeast Candida guilliermondii (J1-JJJ) are reported and its
potential in bioremediation of coastal wetlands is evidenced.
Candida guilliermondii and Saccharomyces cerevisiae
yeasts as inactivated yeasts with lead and cadmium
removal capacity presented similar results in the first
3 h of evaluation.
The native yeast Candida guilliermondii presented
good results as inactivated yeast at 121°C in the removal
of lead and cadmium in the waters from the RCA Wetlands
of Ventanilla under laboratory conditions, so it would be a
yeast with bioremediation potential for this ecosystem.
The isolation of yeasts from groundwater in coastal
wetland ecosystems affected by heavy metals is recom-
mended, in order to explore possible applications in bior-
emediation with native yeasts in affected ecosystems.
Nomenclature
RCA Regional Conservation Area
EQS environmental quality standard
Acknowledgments: The results presented are part of the
doctoral research project sponsored by the National Fund
for Scientific, Technological Development and Technological
Innovation (FONDECYT), currently PROCIENCIA, one of
ten doctoral programs financed by PROCIENCIA in Peru –
Contract 04-2018-FONDECYT/BM. The authors acknowl-
edge and are thankful to the DSR for the technical and
financial support.
Funding information: This research has been financed by
the Concytec-World Bank Project “Improvement and
Expansion of the Services of the National System of
Science, Technology and Technological Innovation”
8682-PE, through its executing unit ProCiencia [Contract
04-2018-FONDECYT/BM], with the aim of contributing to
research in priority lines that includes fragile ecosystems
such as Coastal Wetlands.
Author contributions: Narda Fajardo Vidal – conceptualiza-
tion, formal analysis, funding acquisition, research,
resources, visualization, writing-original draft; Jorge Wong
Dávila – methodology, project administration, supervision,
validation, writing-review and editing.
Conflict of interest: The authors declare no conflict of
interest.
Ethical approval: The conducted research is not related to
either human or animal use.
Data availability statement: All data generated or ana-
lysed during this study are included in this pubblished
article.
 Lead and cadmium removal with native yeast from coastal wetlands
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