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Bioaccumulation of Arsenic in Chlorella vulgaris (Chlorophyta:

Chlorellaceae) in effluent from industrial park Rio Seco and


acute toxicity on Daphnia magna (Crustacea: Daphniidae),
Arequipa, Peru
Ronald E. Huarachi Olivera1; Alex Dueas Gonza1; Ursulo Yapo Pari1 ; Percy Apfata1;
Norbert Tejada Rivera1; Dennis Manuel Cristbal1 & Rosaura Gonzalez Juarez1

Study Area

Problem
Arsenic (As): Metalloid

- Plant of treatment Chilpina,


Arequipa, Peru
Chlorella vulgaris (Chlorophyta:
Chlorellaceae)
713215,93 LO;
162650,76 LS;
2293 m.s.n.m

- Very Toxic Element


- Damages on human health
Environmental Protection Agency (EPA 1986)
- Allowable limit of arsenic (As) in natural waters: 0.05 mg/L
Discharges to the environment
- INDUSTRIAL PARK RO SECO (IPRS): Liquid waste
Big impact on air, water and soil pollution

- INDUSTRIAL PARK RO

SECO (IPRS)
Northeast of the center of the
city of Arequipa; District Cerro
Colorado, Province Arequipa,
Peru.
713559 LO; 162122 LS;
2250 m.s.n.m.

Arsenic in Chlorella: experimental setup


Determination
of
the
capacity of bioaccumulation
of As in industrial effluents
of the IPRS
Gravity flow system

Growth
of
Chlorella
vulgaris
at
different
concentrations of As III
(Na2HAsO3)
T=Negative control
T1= 1.6 mg/L

- 20 mL/min

T2= 3.2 mg/L

- 40 mL/min

T3= 6.4 mg/L

- 80 mL/min

Scope

Microbial bioremediation of arsenic: Interesting option


- High efficiency
- Low-cost
Microbial activity:
- Detoxify
- Mobilize or immobilize the arsenic
Oxidation reduction
Biomethylation
Complexation processes

General Scheme of heavy metal detoxification mechanism mediated by class III metallothineins (MtIII) in microalgae.
Abbreviations: MeL) ,metal complex in solution, (Men+) free heavy metal ion, (X) biotic exocellular ligand, (E) glutamic acid, (C)
cysteine, (G) glycine, (EC) Gammaglutamylcysteine, (ECG)Glutathione, ([EC]2G) Metallothionein n=2, (LMW) low molecular
weight, (HMW) high molecular weigth, (ECS) Gamma glutamylcysteine synthetase, (PCS) phytochelatin synthetase, (HMT1)
vacuolar ABC transporter. * when this step is repeated MtIII of longer chain can be synthesized, MtIII is disassociated when
released to medium (Perales et., al. 2006)

Acute toxicity in Daphnia: experimental


setup
The effluent from the IPRS treated with lower residual As concentration, having gone through
the modified system of flow by gravity.
Acute toxicity of industrial effluents of the IPRS treated with Chlorella vulgaris, in
Daphnia magna.
Count of neonates from 0 to 24 h: LC50 (Median Lethal Concentration)
Negative control
- 250 mL reconstituted water
25% Effluent

Samples were taken every five days for a period of 20 days


Evaluation of counting Chamber of Neubauer

First results
The growth of Chlorella vulgaris according
to concentrations of As III shows an
increase in the number of cells until the 4th
assessment , (6.4 mg/L) 71333.34 org/mL.

The concentration of chlorophyll "a" in a


system
of
flow
by
gravity
with
Chlorella vulgaris shows the presence of
three groups (a, b and c) according to the
test of multiple comparison of Tukey.

System of flow by gravity at different speeds


of flow with Chlorella vulgaris. a) initial
cultivation of 1.5 L of C. vulgaris by tray.
b) effluent from the IPRS treated in the
system of flow by gravity to 20 mL/min.
c) 40 mL/min. d) 80 mL/min.

Shows the presence of two groups (a and


b) according to the test of multiple
comparison of Tukey, the T3 treatment
(6.4 mg/L) which presented higher growth
of Chlorella vulgaris, followed by
treatments T1 and T2, being the control
group which presented lower average
growth.

The residual concentration of industrial


effluents of the IPRS As (Industrial Park Rio
Seco) through a system of flow by gravity with
Chlorella vulgaris shows the presence of two
groups (a and b) according to the Tukey
multiple comparison test.

Samples were taken every


four days for a period of 20
days
- Chlorophyll a
- Residual concentration
of As

- 185 mL reconstituted water


50% Effluent
- 125 mL reconstituted water

- 62.5 mL Effluent PIRS


- 125 mL Effluent PIRS

75% Effluent
- 62.5 mL reconstituted water

- 187.5 mL Effluent PIRS

First results
Industrial effluents of the
IPRS As removal (Industrial
Park Ro Seco) through the
system of flow by gravity
with
Chlorella
vulgaris
shows bioaccumulation of
As effluent to 20 mL/min
with 60.05%, followed by
23.72% decrease retrieved
40 mL/min, while the
smaller
decline
was
obtained to 80 mL/min at
6.94% As present in the
effluents of the IPRS. Also
the model describing the
percent
decrease
As
indicates that if the flow
velocity increases by 1
mL/min As decreases in
0.82%.

Acute toxicity of the IPRS treated with


Chlorella vulgaris, in Daphnia magna
model mathematician Probit shown the
LC50-48h = 3.25 mg/L effluent treated in the
system of flow by gravity to 20 mL/min.

Conclusions
- Maximum growth of Chlorella vulgaris is present in the 4th assessment with
T3 = 6.4 mg/L As III.
- The capacity of bioaccumulation of As with Chlorella vulgaris in effluents of
the IPRS, was measured by indicator of biomass as the concentration of
chlorophyll "a" decreases as speed increases in the system of flow by gravity
with removal of the 60.05% As in 20 mL/min.
- The bioassay of toxicity with Daphnia magna indicates that the (LC50) was
3.25 mg/L at 48 h evaluation.

Contact
Laboratory of Aquatic Biology, Academic Department of Biology,
National University of Saint Augustine (UNSA), Av. Alcides Carrin s/n, Arequipa, Peruvian
E-mail: biologiaacuaticaunsa@gmail.com
*Corresponding author: A. Wolf cel.: 5154958006028 http://ecotoxicologia-bioensayos.blogspot.com