Aquatic Ecosystem Health & Management

ISSN: 1463-4988 (Print) 1539-4077 (Online) Journal homepage: http://www.tandfonline.com/loi/uaem20

Algal growth potential and oxidative stress in

Ankistrodesmus falcatus exposed to waters from
Xochimilco Lake system, México

O. Abeja-Pineda, E. López-López, L. Favari & J. E. Sedeño-Díaz

To cite this article: O. Abeja-Pineda, E. López-López, L. Favari & J. E. Sedeño-Díaz (2015) Algal
growth potential and oxidative stress in Ankistrodesmus falcatus exposed to waters from
Xochimilco Lake system, México, Aquatic Ecosystem Health & Management, 18:2, 221-231

To link to this article: http://dx.doi.org/10.1080/14634988.2015.1040709

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Apr 2015.
Published online: 23 Apr 2015.

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 Algal growth potential and oxidative stress in
Ankistrodesmus falcatus exposed to waters from
Xochimilco Lake system, M
exico
O. Abeja-Pineda,1 E. L
opez,1,* L. Favari,2 and J. E. Sede~no-D
ıaz3
opez-L

1
Lab. de Bioconservaci
on y Manejo, Escuela Nacional de Ciencias Biol
ogicas, I.P.N., Prol. de Carpio y Plan de
Ayala s/n, 11340 M
exico City, D.F., M

exico
2
Departamento de Farmacolog
ıa, Centro de Investigaci
on y de Estudios Avanzados del Instituto Polit
ecnico
Nacional, Apdo. Postal 14-740, 07300 M
exico City, M
exico
3
Coordinaci
on Polit

ecnica para la sustentabilidad IPN. Edif. De la Biblioteca Nacional, 3er Piso, Av. Instituto
Polit

ecnico Nacional s/n, Esq. Wilfrido Massieu, Col. Zacatenco, 07738, M
exico City, D.F., M

exico
*Corresponding author: eulopez@ipn.mx

The aim of this study was to assess the toxic effect of the water of Xochimilco Lake on a native
microalgae species of the first trophic level, Ankistrodesmus falcatus. We evaluated oxidative stress
(lipid peroxidation, and antioxidant activity of the enzymes: superoxide dismutase, catalase and
glutathione peroxidase), and algal growth of A. falcatus, when exposed in bioassays to waters from
three zones of Xochimilco Lake (urban, tourist, and agricultural), in four study periods; the responses
were compared with those of a batch control. Water quality shows serious deterioration in the
Xochimilco Lake system. Biomonitoring with biomarker responses provided information about the
spatio-temporal impact of pollution on A. falcatus. Water samples of the three study zones of Xochimilco
Lake induced oxidative stress in A. falcatus, and impaired its antioxidant enzymes and algal growth
responses. Alteration detected in algal growth and oxidative stress of A. falcatus when exposed to
waters from all sites and study periods make evident the toxic effect of the water from the Xochimilco
Lake system.

Keywords: algal growth impairment, biomonitoring, wetland, urban lake, algae bioassay, bio-
markers, lipid peroxidation, algae inhibition/stimulation

Introduction deterioration (Ochoa and Gonz
alez, 2008), and

Pollution of inland waters by industrial and
domestic wastewaters, without adequate treatment,
generates complex mixtures of organic and inor-
ganic xenobiotics provoking water quality
can cause imbalances in the different metabolic
processes of aquatic biota (Sherry, 2003). Xochi-
milco Lake system (XLS), located at the Mexico
City, has had an extensive historical and cultural
tradition. In 1987, the UNESCO designated

Color versions of one or more of the figures in the article can be found online at www.tandfonline.com/uaem.

221
Aquatic Ecosystem Health & Management, 18(2):221–231, 2015. Copyright Ó 2015 AEHMS. ISSN: 1463-4988 print / 1539-4077 online
DOI: 10.1080/14634988.2015.1040709
222 Abeja-Pineda et al. / Aquatic Ecosystem Health and Management 18 (2015) 221–231
Xochimilco as a “world historical and cultural
heritage.” Biodiversity of XLS is valuable, since it
is one of the last vestiges of the lakes of the Basin
of Mexico City (Ezcurra, 1990), some of its native
species have significant biological and commercial
importance; furthermore many migratory birds
temporarily increase its diversity contributing to
its ecological uniqueness (Mart
ınez-Cruz et al.,
2006). In 2004, XLS was added to the Ramsar
List of Wetlands of International Importance
(RCW, 2008). Nowadays, XLS faces various prob-
lems: urban sprawl, overexploitation of aquifers,
drying of springs, and discharge of wastewater
from two treatment plants that replaced the origi-
nal springs that supplied the lake. Additional water
inputs include domestic wastewater from untreated
irregular human settlements; wastes from livestock
barns, from tourism activities, and pesticide resi-
dues (Boj
orquez and Amaro, 2003). Some xenobi-
otics promote oxidative stress in organisms, when
they undergo oxidation-reduction transformation
into reactive oxygen species (ROS), such as the
superoxide radical, hydrogen peroxide, or hydroxyl
radical (Atli and Canli, 2010). These ROS can
cause deleterious effects at different levels of bio-
logical organization (e.g. molecular, biochemical,
and physiological), that can be detected with early
warning biomarkers that identify changes at either
biochemical or cellular levels (Sherry, 2003).
Lipid peroxidation (LPO) is one of the main
effects of ROS, and has been shown to occur from
the toxic action of environmental pollutants, lead-
ing to loss of cellular function (Storey, 1996). In
healthy cells, ROS are detoxified by antioxidant
defense systems such as antioxidant enzymes:
superoxide dismutase (SOD), catalase (CAT), and
glutathione peroxidase (GPx), that inhibit the for-
mation of pro-oxidant agents, therefore, these
enzymes may be used to detect sensitive signals
for deleterious effects of environmental pollution
(Van der Oost et al., 2003).
The aim of this study is to assess the toxic
effect of the water of Xochimilco Lake in the
microalgae Ankistrodesmus falcatus (a native spe-
cies of XLS). The algal growth potential and oxi-
dative stress was assessed from exposure of A.
falcatus to three contrasting conditions from
Xochimilco Lake in four periods of study. Bio-
markers analyzed were: LPO, SOD, CAT and
GPx, and the growth inhibition or stimulation of
the exposed microalgae were tested and contrasted
to a batch control. Marker responses were
analyzed with an Integrated Biomarker Response
(IBR) and related to water quality parameters.
Material and methods
Study area
Xochimilco Lake has an approximate area of
423 km2 at an altitude of 2240 masl, the zone has
a dry season from November to April and a rainy
season from May to October. The study sites were
located in the three zones that are distinguished in
the XLS by the land use (Figure 1): The urban
zone (UZ 19 16.337´N-99 06.038´W) character-
ized by irregular settlements and domestic waste-
water discharges, the tourist zone (TZ 19 17.055´
N-99 06.022´W), where tours are offered in boats
“trajineras,” and the agricultural zone or
“chinampas” (AZ 19 16.631´N-99 05.620´W)
with cultivation of vegetables, ornamental plants,
and also with activities of cattle farms.
Four sampling periods were analyzed: the rainy
season (May and August 2009) and the dry season
(November 2009 and February 2010). At each
study site, several parameters were recorded in situ,
using a multiparameter probe Quanta: dissolved
oxygen (DO mg l¡1), conductivity (mS cm¡1),
pH and temperature ( C). For each site and period
of study, water samples of 500 ml for water quality
analysis were collected in duplicate. Also, water
samples for bioassays were collected.
Water quality analysis
Variables determined for water quality were:
nitrites (NO2 mg l¡1), nitrate (NO3 mg l¡1),
ammonia (NH3 mg l¡1), total nitrogen (NT
mg l¡1), orthophosphates (PO4 mg l¡1), total phos-
phorus (PT mg l¡1), sulfate (SO4 mg l¡1) and color
(Pt-Co units) using the techniques of DLR/2500
Figure 1. Location of study sites in the Xochimilco Lake
system.
 Abeja-Pineda et al. / Aquatic Ecosystem Health and Management 18 (2015) 221–231
HACH spectrophotometer. Hardness (CaCO3 mg
l¡1), chloride (Cl mg l¡1), alkalinity (mg l¡1), bio-
chemical oxygen demand (BOD5, mg l¡1), were
analyzed following American Public Health Asso-
ciation (APHA) (2005). Total and fecal coliforms
were analyzed by the most probable number
method (MPN) according to APHA (2005). Con-
centration of metals: Pb (mg l¡1), Cd (mg l¡1), Zn
(mg l¡1) and the metalloid As (mg l¡1) in water
from each study sites and sampling periods were
analyzed after an acid digestion by the method of
flame atomic absorption spectrophotometer (GBC
Avanta Version 2.01), as indicated by the Mexican
guidelines NMX-AA-051-SCFI-2001.
Bioassays with Ankistrodesmus falcatus
for algal growth potential and for
biochemical tests
The strain of A. falcatus was provided by Cen-
tre for Research and Advanced Studies of Instituto
Polit
ecnico Nacional (Lab. 16). The algae were
grown in medium BG-11 which contains: NaNO3
(17.6 mM l¡1), K2HPO4 (mM l¡1), MgSO4
(0.03 mM l¡1), CaCl2 (0.24 mM l¡1), citric acid
(0.031 mM l¡1), ferric ammonium (0.021 mM
l¡1), Na2 EDTA (0.0027 mM l¡1), Na2CO3
(0.19 mM l¡1), algae were previously rinsed with
NaHCO3 to remove the culture medium according
to the procedure of APHA (2005). For the bioas-
say, inoculates of 50 ml of previously rinsed algae
containing 19,600 cells ml¡1 (§784) were spiked
in glass tubes with 10 ml of water from the study
sites (UZ, TZ and AZ) each assay with 6 replicates
(n D 6), water samples were previously filtered
through a 0.40 mm membrane and sterilized in an
autoclave (15 psi, 121 C, for 15 min) (L
opez-
L
opez and D
avalos-Lind, 1998). Cultures were
maintained at room temperature 21 § 0.5 C,
12:12 h light:darkness for 96 h, illuminated by
fluorescent lamps: cold light with light intensity of
1000 lux §10%, the cultures were stirred before
reading the absorbance. Due that the XLS is a
very impacted environment by different anthropo-
genic activities there is not possible to use a refer-
ence site, in consequence we use a laboratory
control. Similarly bioassays were mounted in a
control batch (n D 6 replicates) with 10 ml of cul-
ture medium BG-11. Absorbance readings were
taken (for each of the replicates) from the start of
the assay and every 24 h until the end of the
223
bioassay (72 h), using a Hach spectrophotometer
DRL 2500 at a wavelength of 620 nm. A similar
set of bioassays were set up (n D 6 replicates),
according to the procedure outlined above for bio-
chemical assays. At the end of the bioassay (72 h)
algae were centrifuged at 2000 rpm for 15 min,
the supernatant was decanted and the cell packs
were immersed in ice to prepare homogenates
using an electric homogenizer (Biospecific prod-
uct, Inc. Model 398). Three replicates of each bio-
assay were used to prepare 10% homogenates of
the cell packs of A. falcatus with 0.1 M potassium
phosphate buffer (pH D 7.4) to assess LPO and
CAT activity. The remaining three replicates of
each bioassay were used to prepare 10% homoge-
nates of the cell pack of A. falcatus with 50 mM
potassium phosphate buffer (pH D 7.4) to assess
SOD and GPx activities.
Inhibition/stimulation of algal
growth potential
The daily growth rate (mday¡1) was calculated
according to NAERP and EPA (1971): m D Ln (F2/
F1)/t2-t1, where t1 is the start time and t2 is the time at
the end of the bioassay (72 h), F1 and F2 is the absor-
bance at t1 and t2, respectively. The inhibition/stimu-
lation of algal growth potential was calculated based
on the specific algal growth rate (m) of the test sam-
ple and control, following the equation proposed in
ISO 8692 (2004): Imi D ((mc-mi)/mc)*100, where:
Imi is the percentage inhibition for the test study site
i; mi is the average growth rate for the study site i; mc
is the average growth rate for the batch control. A
negative value of Imi means growth stimulation; a
positive value means growth inhibition.
Biochemical tests
All biochemical assays were evaluated in dupli-
cate and readings were done with a Spectronic
20D spectrophotometer. All biomarkers were
determined in aliquots of the A. falcatus homoge-
nates at 10%. The LPO was determined following
Buege and Aust (1978). For CAT activity, H2O2
consumption was measured at 480 nm (Cohen
et al., 1970). CAT activity was calculated as the
first-order reaction rate constant of the H2O2
decomposition. SOD activity was determined fol-
lowing Sun et al. (1988). The GPx activity was
assessed following Lawrence and Burk (1976)
224 Abeja-Pineda et al. / Aquatic Ecosystem Health and Management 18 (2015) 221–231
with cumene hydroperoxide as substrate. The
enzyme activity was quantified by measuring the
oxidation of NADPH at 1 min. intervals for
4 min., and the enzyme activity was calculated as
nmol NADPH oxidized min¡1mg¡1 of protein.
The protein content in all the samples was esti-
mated by the method of Bradford (1976) using
bovine serum albumin as standard.
Statistical analyses
Analysis of variance (ANOVA) with each
group of variables: water quality, were performed
to determine significant differences between sites
according to the parameters analyzed, a posteriori
Tukey tests were performed when necessary to
distinguish significant differences between sites,
the significance was considered at p < 0.05. For
algal growth assays mean values (n D 6) of the%
inhibition/stimulation was assessed for each site
and study period, significant differences among
control and exposure cultures for each site and
study period were evaluated by one-way ANOVA
and Tukey test (p < 0.05).
Water quality variables, biomarkers and inhibi-
tion/stimulation of algal growth potential were
analyzed by a redundancy analysis (RDA), which
identified those variables that are related to the
responses of A. falcatus. Two matrices were used:
one with water quality parameters and metals and
the second with biomarker responses and inhibi-
tion/stimulation algal growth by site and sampling
periods. These variables were previously trans-
formed (Y D log (xC1)). Statistical tests were per-
formed using the software XLSTAT-Pro 2010.
The IBR was assessed with values of the bio-
markers which were normalized and standardized
by the method proposed by Beliaeff and Burgeot
(2007), which allows analysis of multiple bio-
marker data by using star graphs. The magnitude
of the response of each biomarker is represented
as a vector in the star graph and joining the ends of
the vectors the enclosed area represents a numeri-
cal value of the IBR.
Results
Water quality
Water characteristics reflect XLS is highly min-
eralized, and fertilized; conductivity showed
higher values (mean values 662 to 920 mS cm¡1),
the water is highly alkaline (137 to 288 mg l¡1),
rich in CaCO3 (69 to 120 mg l¡1) and SO4 (38 to
67.5 mg l¡1). DO showed the lowest values during
May and August, and the higher were in TZ
throughout the sampling period. Higher BOD5,
were observed in AZ and UZ in May and Febru-
ary. The highest alkalinity, CaCO3, Cl and con-
ductivity were recorded in November and
February (p < 0.05), and pH ranged from 7.9 to
9.5. NO3 and NO2 were higher in the UZ in
August, November and February (p < 0.05), and
NH3 showed the highest concentrations in May at
all study sites (p < 0.05). Orthophosphates were
higher in the UZ and AZ (p < 0.05). Total and
fecal coliforms were greater in UZ and TZ (p <
0.05). Pb was detected at all sites and periods of
study. Cd was detected in almost all the sites and
periods of study except in AZ and UZ in Novem-
ber, higher values were detected in all the study
sites in May, in UZ and TZ in August, and the TZ
in November and February. Zn showed higher
concentrations in the AZ in all the study periods,
in UZ during May, and in TZ in August. As was
detected in the three study sites during November,
in February As was detected in the TZ and AZ
(Table 1).
Inhibition/stimulation of algal
growth potential
Bioassays in the three study sites showed algal
growth inhibition (AGI) in at least one period of
study: in UZ and TZ in August (rainy season), the
inhibition by site was ordered as follows: UZ >
TZ (Figure 2). In November (cold dry period)
AGI occurred in the TZ and AZ sites (Figure 2).
In May (rainy season) and February (dry season)
algal growth stimulation was detected in the three
study sites (Figure 2).
Redundancy analysis and integrated
biomarker response
The RDA showed segregation of the sites and
periods of study and their relationship with the
environmental variables and biomarkers assessed
in A. falcatus. The first two components reached
94.50% of the explained variance. In the right
quadrants of Figure 2a, sites UZ, TZ and AZ of
 Table 1. Physical and chemical parameters of Xochimilco Lake system water quality in the three study sites over an annual cycle.

Urban zone Tourist zone Agricultural zone

2009-2010 2009-2010 2009-2010

May August November February May August November February May August November February

¡1
Conductivity (mS cm ) 786 713 903 852 785 662 910 860 810 710 920 872
Chlorides (mg l¡1) 56.98 §2.82ab 55.78 §0.28c 76.17 §0.84a 78.57 §0.28b 63.98 §1.4 57.58 §0.56a 72.97 §2.5 83.77 §1.97 59.18 57.38 §0.28a 77.37 §0.28 79.77 §0.28
SS (mg l¡1) 33 22 19.5 48.5 102.5 49 37.5 34.5 196 29 36 64
Color (Pt-Co units) 61 56 79 52.5 110 49 85 32.5 127 59 81.5 50.5
Hardness (mg l¡1) 69 §0.014c 80 §0.028b 120 §0.28a 105 §0.07a 64 §0 70 §0 93 §0.04b 111 §0.01a 76.5 §0.04a 76 §0a 97 §0.01b 99 §0.01b
Sulfates (mg l¡1) 53.5 59 49 53.5 56 65.5 38 § 1.41a 60 60 63.5 57.5a 67.5
pH 8.5 8.5 5.6 7.9 8.4 8.7 7.9 9.5 8.6 8.6 8 9.2
Alkalinity (mg l¡1) 184.4 §0c 137.6 §0b 255.6 §0.04a 260.4 §0.14a 188 §0a 184 §0.05a 278 §0.042b 255.2 §0c 212 §0.14d 172.8 §0.14c 288.4 §0.14a 261.2 §0.042b
Nitrites (mg l¡1) 0.314 0.124 0.159 0.236 0.077 0.051 0.12 0.109 0.049 0.066 0.056 0.052
Nitrates (mg l¡1) 2.95 §0.07a 4.15 §0.07 4.05 §0.07 3.95 §0.07 1.5 §0.14a 1.7 §0.28a 3.9 §0c 2.4 §0b 1.8 §0 3.65 §0.07a 2.05 §0.07 1.75 §0.07
Ammonia (mg l¡1) 2.36 1.11 0.99 0.72 1.51 0.905 0.79 0.92 2.01 0.62 0.78 0.83
Orthophosphates 6.16 5.93 5.71 6.22 5.16 3.11 6.65 6.04 5.6 5.48 6.44 6.42
(mg l¡1)
Total Coliforms 1600 500 1600 1600 1600 280 1600 1600 1600 280 1600 350
(MPN 100 ml¡1)
Fecal Coliforms 350 34 280 33 23 9 2 14 4 7 2 11
(MPN 100 ml¡1)
DO (mg l¡1) 3.5 2.81 7.63 9.69 8.53 8.49 10 9.23 5.42 8.37 11.82 7.51
BOD5 (mg l¡1) 26.56 18.73 11.24 22.82 24.52 22.82 14.98 8.85 24.18 10.21 21.45 49.87
NT (mg l¡1) 13.4 §0.42b 32.5 §4.94a 13.1 §0.42b 37 §1.41a 9.45 §0.49 25 §1.41b 9.7 §0.14 33 §1.41a 14.7 §0.84c 28.5 §0.70b 8 §1.41d 55 §7.07a
PT (mg l¡1) 8.44 §0.11b 7.19 §0.57 6.77 §0.09 10.02 §0.02a 8.31 §0.26 4.17 §0.01a 8.38 §0.36 8.2 §0.19 8.51 §0.57b 7.47 §0.07c 9.51 §.04ab 10.36 §0.19a
Temperature ( C) 22.52 21.55 16.41 19.58 23.08 22.11 16.8 18.12 20.72 22.01 15.59 17.52
Pb (mg l¡1) 0.4 0.31 0.4 0.4 0.36 0.31 0.38 0.33 0.32 0.33 0.38 0.43
Cd (mg l¡1) 1.16 1.61 0.09 — 1.14 0.05 2.4 1.55 1.07 1.16 — 0.38
Zn (mg l¡1) 9.4 — — — — 4.8 — — 4.6 4.4 4.2 2.6
As (mg l¡1) — — 0.06 — — — 0.09 0.62 — — 0.08 1.37

-: Non detectable. Different letters (a, b, c, d) denote significant differences between periods of study for the same study site (p < 0.05).

225
226 Abeja-Pineda et al. / Aquatic Ecosystem Health and Management 18 (2015) 221–231
Figure 2. Percentage inhibition/stimulation of algal growth (A. falcatus) at 72 h. (a) May, (b) August, (c) November 2009 and (d)
February 2010. UZ: Urban zone, TZ: touristic zone and AZ: agriculture zone. Bars show error standard. *Significant differences (p
< 0.05) compared to control.
May (rainy season), TZ of August (wet season),
and TZ of November (cold dry season), are associ-
ated with the highest values of pluvial precipita-
tion, NO3, NO2, NH3 and high color scores (Pt-
Co). In the upper left quadrant of the biplot, the
UZ in August, November and February, are asso-
ciated with the highest concentration of total coli-
forms, suspended solids and BOD5. In the left
lower quadrant of the biplot, the TZ of February
and AZ of February and November are highly
related with the highest values of DO, pH, SO4,
total dissolved solids, orthophosphates, Cl, alka-
linity, CaCO3, Total N, Total P and conductivity
(Figure 3a).
The RDA showed A. falcatus exposed to waters
of XLS exhibited the higher LPO levels in the AZ
in August (wet season) and the UZ in May, sites
with the highest concentration of As. A higher
enzymatic activity of GPx and CAT was detected
in the AZ, TZ and UZ in May; particularly a high
CAT activity was detected in AZ, and TZ of
August and TZ in November (Figure 3b). The
high activity of GPx was related to the highest
concentration of Cd in water of the study sites.
Furthermore, it was observed a higher growth rate
of A. falcatus in TZ in November (Figure 3b).
Finally, in the lower left quadrant are located TZ
and AZ in February and AZ in November that are
highly related with the highest Pb and Zn concen-
tration in water and the lowest growth rate of A.
falcatus (Figure 3b).
The IBR of A. falcatus by study site, showed
that in May the UZ reached the highest value
(IBR D 10.06) in concurrence with higher GPx
and CAT. In August the highest IBR was in the
AZ, when LPO, GPx and CAT had the major con-
tribution to this response, while SOD decreased.
In the UZ and TZ of August the SOD was highly
depleted. The IBR in November was higher in the
UZ, all biomarkers contributed to IBR. In Novem-
ber LPO decreased in UZ, when compared to
May, but the antioxidant enzymes were increased.
Likewise, in the AZ, SOD activity decreased, but
the greater antioxidant enzyme response was by
GPx and CAT, however, there was an increase in
LPO. In February, the highest IBR was in the UZ,
all biomarkers contributed to the IBR. In the TZ,
the IBR was determined by GPx, CAT and LPO
while SOD was depleted. The AZ displayed a
decreased GPx; however, CAT and LPO increased
(Figure 4). The IBR analyzed by sampling periods
(which considers the summary of the IBRs in all
the study sites for each study period), showed the
following order: November > February > May >
 Abeja-Pineda et al. / Aquatic Ecosystem Health and Management 18 (2015) 221–231 227

Figure 3. Biplots of the redundancy analysis showing: (a) relationships between study sites and environmental factors. Labels are
as follows: Solid squares (&) represent study sites in different periods of study: urban zone (UZ), touristic zone (TZ), agriculture
zone (AZ), the letter after the study site represents the month: May (M), August (A), November (N), February (F); and the vectors
ending with solid circles () represent environmental variables: color (Pt-Co), nitrate (NO3), nitrite (NO2), ammonia (NH3), alkalin-
ity (ALK), hardness (CaCO3), chloride (Cl), conductivity (COND), Temperature (T), precipitation (pp), orthophosphates (PO4),
total coliforms (Col-T), (pH), biochemical oxygen demand (BOD), dissolved oxygen (DO). (b) Relationship between study sites
and responses of A. falcatus in bioassays. Labels are as follows: Solid squares (&) represent study sites in different periods of study
as in (a); the vectors ending with solid circles () represent the heavy metals and metalloids: cadmium (Cd), lead (Pb), zinc (Zn),
arsenic (As); and the solid triangles (~) represent responses in A. falcatus: growth rate (m), level of lipid peroxidation (LPO),
superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx).

August, whereas the IBR by study sites (which
consider the summary of the IBRs in all study peri-
ods for each study site) showed the following
order UZ > AZ > TZ (Figure 4).
Discussion
Physicochemical parameters of XLS reflect
poor water quality. Conductivity, alkalinity, Cl
and CaCO3 showed higher values. These condi-
tions likely resulted from: water treatment facility
discharge, which is the main source of water to
XLS; and untreated wastewater discharges from
the UZ. The high concentrations of N and P are
likely caused by land use that primarily consists
of agriculture, cattle raising, housing, flower nurs-
eries, and recreation. Higher values of NO3, NO2
and NH3 were detected in the UZ particularly
during the rainy season, where discharge of
domestic wastewater is more frequent. Surface
water inflow is often the major input of nutrients
in several wetlands, with higher inputs during
rainy seasons (Sutula et al., 2001). Given that
concentrations of NT, PT, NO3, NO2, NH3, and
PO4 are all high in XLS, it is evident their highly
eutrophication process. Coliforms were present
at all periods and study sites. The highest coli-
forms densities were present in the UZ and TZ
that are strongly impacted by domestic dis-
charges. Fecal coliform (MPN) in XLS does not
exceed NOM-003-ECOL (1997) guidelines for
the reuse of wastewaters treated for public serv-
ices, however, coliforms are indicators of patho-
genic pollution representing a sanitary risk in
Xochimilco Lake.
The DO was at acceptable levels (mean
7.76 mg l¡1), capable of supporting resident
aquatic biota (Radwan et al., 2003); however, UZ
had the lowest values, 2.8 and 3.5 mg l¡1 in
August (rainy) and May (rainy season) respec-
tively, which are likely related to the organic mat-
ter that increased in those periods.
Among the metals assessed Pb and Zn and the
metalloid As in all periods and study sites did not
exceed the Mexican guidelines by the NOM-001-
ECOL-1996. Contrary, Cd concentrations in all
study in May, UZ and AZ in August and TZ
in November and February exceeded those
guidelines.
228 Abeja-Pineda et al. / Aquatic Ecosystem Health and Management 18 (2015) 221–231

Figure 4. Integrated biomarker response (IBR) in A. falcatus for study sites; urban zone (UZ), touristic zone (TZ), agriculture zone
(AZ); and periods of study: May, August, November, February. Level of lipid peroxidation (LPO) catalase (CAT), superoxide dis-
mutase (SOD), glutathione peroxidase (GPx). IBR CONT: Control of IBR in A. falcatus.
Although we detected high nutrient concentra- effects on algal growth in August and TZ in
tions in XLS, algal growth inhibition was observed November. Previous studies in XLS identified the
in the UZ in August and in AZ in November (Fig- presence of some heavy metals, such as Cr, Hg,
ures 1b and c), showing that UZ exerts negative Cd, Pb, Zn, and the metalloid As (Boj
orquez and
 Abeja-Pineda et al. / Aquatic Ecosystem Health and Management 18 (2015) 221–231
Amaro, 2003), in this study we assessed Cd, Pb,
Zn, and As. Cd can induce a wide spectrum of
toxic effects on the enzymatic activities of plants:
it can either bind to functional groups (e.g. sulfhy-
dryl, carboxyl, imidazole) or replace essential ele-
ments that participate at the active sites of
enzymes, both resulting in inhibition of photosyn-
thesis, respiration, biosynthesis of photosynthetic
pigments, and cell growth (Aravind and Prasad,
2004). Metals such as Cu and Zn, are essential to
photosynthetic organisms, but not at high concen-
trations that impair photosynthesis (Li et al.,
2006). Prasad and Prasad (1981) found that Pb at
higher concentrations provoked lethality in A. fal-
catus. However, a combination Cd-Pb provokes
antagonistic effects: toxicity is lower in assays
with combined Cd-Pb when compared to their
individual effects. For As, Pawlik-Skowronska
et al. (2004) found that accumulation and toxicity
in algae was related to pH. In XLS algal growth
stimulation was detected in UZ and TZ in August
and in the TZ and AZ in November (during the
rainy and post- rainy seasons). Stimulation of
algae growth has been detected in the presence of
vegetable tannin water extracts, hormones, and
some fungicides and pesticides, all which exert
induction at low doses and inhibition at high doses
(i.e. hormetic effect) (De Nicola et al., 2007; Ceder-
green and Streibig, 2007). However, higher concen-
trations of the same organic pollutants will
irreversibly damage algal cells, and growth is
completely inhibited (Gong et al., 2009). These pol-
lutants, at high concentration, can also produce large
amounts of ROS in algal cells and, thereby, lead to
membrane lipid peroxidation, cell damage, and inhi-
bition of cell growth (Kamaya et al., 2006). In XLS
high nutrient concentrations and some organic com-
pounds could be entering in the rainy season provok-
ing the stimulation of algal growth.
Enzymatic antioxidants can scavenge free radi-
cals, catalyzing defense reactions that inhibit the
oxidative damage caused by ROS (Elbaz et al.,
2010). Some researchers (De Nicola et al., 2007;
Cedergreen and Streibig, 2007), have considered
that hormesis may be related to oxidative stress
and ROS (hydrogen peroxide, superoxide anions,
and hydroxyl radicals). To prevent damage to cel-
lular lipids, proteins, and DNA, the antioxidant
system is activated to defend and repair unfavor-
able conditions (Sureda et al., 2008). A decrease
in SOD can cause excessive production of ROS in
algal cells, resulting in membrane LPO and cell
229
damage. Atli and Canli (2010) found excessive
production of superoxide radicals causes oxidation
of cysteine, which deactivates SOD. We detected
that stimulation in algal growth was also related
with the periods and sites with highest decreases
in SOD (UZ and TZ in August and TZ and AZ in
November) (Figure 3). An increase in CAT activ-
ity in A. falcatus is related to its antioxidant
defense against increased LPO, which is generated
from superoxide and hydrogen peroxide. Atli and
Canli (2010) found this enzyme compensated for a
decrease in antioxidant enzymes, such as SOD and
GPx. Similarly, Pinto et al. (2003) found GPx to
be one of the most important components in cellu-
lar antioxidant defense systems, as it reduces
hydrogen peroxide to water and alcohol. In this
study, the increased activity of GPx in A. falcatus
was likely due to the presence of hydrogen perox-
ides that increased LPO in the algae, reflecting the
protective function of algae in the detoxification
and excretion of free radicals. In addition,
although CAT and GPx increased, presumably to
help algal cells eliminate ROS and protect them-
selves, the damage to active oxygen scavengers,
like SOD, disabled proper elimination of excessive
active oxygen, causing damage to other cell func-
tions rather than provoking inhibition in algal
growth. In contrast, the high availability of
nutrients, particularly nitrogen, increases both the
ability of algae to accumulate toxic metals and the
production of ROS that oxidize proteins, lipids,
and nucleic acids, all causing changes in cell struc-
ture by oxidative stress (Pokora and Tukaj, 2010).
Conclusions
In our assays, we found higher LPO as com-
pared to the batch control, which is an indicator of
toxicity of the XLS. The IBR assessment showed
the integration of oxidative stress biomarkers dis-
playing that UZ, TZ, and AZ had IBR peaks in
May, November, and August, respectively. How-
ever, during the whole sampling period, the pattern
was similar: increased LPO provoked increased
activity of GPx and CAT, but, in all cases, the
decrease in SOD was evident, suggesting damage
in this antioxidant enzymatic system.
Acknowledgements
Authors thank Ma. Teresa Garc
ıa for technical
assistance.
230 Abeja-Pineda et al. / Aquatic Ecosystem Health and Management 18 (2015) 221–231
Funding
This research was financed by the Institute of
Science and Technology of the Federal District,
M
exico project number ICyT No-86.
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