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An assessment of chemical and physical parameters, several contaminants

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DOI: 10.1080/10934520903388517 · Source: PubMed

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LESA #439029, VOL 45, ISS 1

An assessment of chemical and physical parameters, several

contaminants including metals, and toxicity in the seven
major wastewater treatment plants in the state
of Aguascalientes, Mexico
FÉLIX TORRES GUZMÁN, FRANCISCO JAVIER AVELAR GONZÁLEZ and ROBERTO RICO
MARTÍNEZ

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An assessment of chemical and physical parameters, several contaminants including metals, and toxicity in the seven major
wastewater treatment plants in the state of Aguascalientes, Mexico
Félix Torres Guzmán, Francisco Javier Avelar González and Roberto Rico Martı́nez
 Journal of Environmental Science and Health Part A (2010) 45, 1–13
Copyright
C Taylor & Francis Group, LLC

ISSN: 1093-4529 (Print); 1532-4117 (Online)

DOI: 10.1080/10934520903388517

An assessment of chemical and physical parameters, several

contaminants including metals, and toxicity in the seven
major wastewater treatment plants in the state
of Aguascalientes, Mexico

5 FÉLIX TORRES GUZMÁN1 , FRANCISCO JAVIER AVELAR GONZÁLEZ2 and ROBERTO RICO
MARTÍNEZ3
1
Departamento de Quı́mica, Universidad Autónoma de Guerrero, Guerrero, México
2
Departamento de Fisiologı́a y Farmacologı́a, Centro de Ciencias Básicas, Universidad Autónoma de Aguascalientes,
Aguascalientes, México
3
10 Departamento de Quı́mica, Centro de Ciencias Básicas, Universidad Autónoma de Aguascalientes, Aguascalientes, México

Forty-eight hours acute toxicity tests were employed with Daphnia magna and Lecane quadridentata to assess the influents and
effluents of the seven most important wastewater treatment plants (WTP) in the state of Aguascalientes, Mexico, during the 2006 dry
and rainy seasons. The WTP of the City of Aguascalientes treated 1948 Ls−1 . The remaining six plants treated wastewater in the range
from 28 to 93 Ls−1 . Plants efficiently removed toxicity when Daphnia magna was used as a model organism, but performed poorly
15 when the freshwater rotifer Lecane quadridentata was employed. It was observed that biochemical oxygen demand (BOD5 ), chemical
oxygen demand (COD), conductivity, dissolved oxygen (DO), pH, phenol, methylene blue active substances (MBAS), temperature,
and total suspended solids (TSS) were within the maximum allowed levels (MAL) in the effluents during the rainy season. Whereas
the BOD5 , total nitrogen, total dissolved solids (TDS) and TSS showed levels greater than the MAL in effluents during the dry
season. The levels of BOD, MBAS, and total nitrogen were greater than the MAL in influents. In contrast, the values of TDS and
20 TSS in influents were above the MAL during the rainy season. In the dry season the levels of aluminum (Al), iron (Fe), zinc (Zn),
manganese (Mn), chromium (Cr), mercury (Hg) and lead (Pb) in influents and Fe, Mn and Pb in effluents were above the MAL.
During the rainy season the levels of Mn, Cr and Hg in influents and those of Mn in effluents were above the MAL. When D. magna
was used as model organism, acute toxicity units (aTU) for influents ranged from 0.2 to 4.4 and from 0.1 to 0.2 for effluents. These
values for effluents are acceptable according to international guidelines. However, when L. quadridentata was the model organism,
25 ranges were from 2.0 to 8.3 aTU in influents and from 1.6 to 2.6 aTU in effluents. The treated water that discharges into the San
Pedro River would be considered toxic. The results of the toxicity tests demonstrated that the freshwater rotifer Lecane quadridentata
was more sensitive (in 89.3% of the samples) than the cladoceran Daphnia magna when both organisms were exposed to the influents
and effluents of the wastewater treatment plants.
Keywords: Metal toxicity, acute toxicity tests, LC50, Daphnia magna, Lecane quadridentata.

30 Introduction
The state of Aguascalientes, Mexico ranges from 22◦ 27 to
21◦ 38 N and from 101◦ 53 to 102◦ 52 W[1] . The San Pedro
River is the most important river in the state, and belongs
to the Lerma-Santiago Hydrological Region. The effluents
35 of the main wastewater treatment plants discharge to the
Address correspondence to Félix Torres Guzmán. Avenida
Lázaro Cárdenas s/n, ciudad universitaria. Chilpancingo,
Guerrero, México. C. P. 39660. E-mail: torresfelix2000@yahoo.
com.mx
San Pedro River. The river also catches the municipal and
industrial untreated discharges and runoff.[1,2]
Treated water is used to irrigate agricultural fields and
green areas. Given the scarcity of water, it is necessary to
monitor this treated wastewater and to make sure it con- 40
forms to the required quality for these activities.[3] However,
the quality of the effluents cannot be based solely in phys-
ical and chemical parameters.[4,5] Toxicity tests represent
the best tool to assess the toxicity of a complex mixture
of toxicants typically found in wastewater.[5] The aquatic 45
toxicity tests considered by Mexican legislation use model
organisms: Daphnia magna[6] , Artemia franciscana[7] and
 2 Guzmán et al.

Fotobacterium fosforeum.[8] However, the freshwater rotifer
Lecane quadridentata has demonstrated higher sensitivity
50 to lower concentrations of toxicants found in wastewater
than D. magna.[9]
Nevertheless, due to the variety and complexity of the
contaminants mixed in the wastewater, it is difficult to
discover the real cause of the toxicity shown by a par-
55 ticular discharge on aquatic organisms.[4] However, there
are some strategies like Toxicity Identification Evaluation
(TIE) and Whole Effluent Toxicity (WET) that use physico-
chemical and biological criteria to fractionate and analyze
the whole effluent using toxicity tests on each fraction, al-
60 lowing the identification of the major toxicants involved
in the generation of toxicity. Such approaches determine if
a particular discharge is potentially toxic to the biota in-
habiting a reservoir.[10,11] These analytical approaches have
become valuable tools to assess the environmental impact
65 of discharges,[12,13] allowing identification of the chemical
substances responsible for most of the observed toxicity.[14]
The goal of this study is to assess the physical and chem-
ical parameters, levels of selected contaminants (including
some metals), and levels of toxicity of both influents and
effluents of the seven most important wastewater treatment 70
plants in the state of Aguascalientes, Mexico during an an-
nual hydrological cycle.
Materials and methods
Collection of samples from the wastewater treatment plants
Samples were taken from influents and effluents of the seven 75
most important wastewater treatment plants (Fig. 1) lo-
cated in six municipalities of the state of Aguascalientes
during the 2006 dry (February and March) and rainy sea-
son (October and November). We selected the dry and rainy
season because they represent the most important stages of 80
the hydrological cycle in Aguascalientes.[15] The wastew-
ater treatment plants were selected because they treated

Fig. 1. Localization of seven water treatment plants investigated in Aguascalientes, Mexico. 1. AGS = City of Aguascalientes. 2. LS
= Los Sauces, 3. PIVA = Parque Industrial del Valle de Aguascalientes. 4. JM = Jesús Marı́a. 5. SFR = San Francisco de los Romo.
6. PA = Pabellón de Arteaga. 7. RR = Rincón de Romos.
 Wastewater in seven treatment plants in Aguascalientes, Mexico
the greatest volumes (Ls−1 ) in the state: City of Aguas-
calientes (AGS) 1948.0, Los Sauces (LS) 93.76, Rincón de
85 Romos (RR) 85.22, Pabellón de Arteaga (PA) 59.08, Jesús
Marı́a (JM) 52.60, San Francisco de los Romo (SFR) 36.21,
and Parque Industrial del Valle de Aguascalientes (PIVA)
28.78. Samples were collected in 5-L polypropylene bottles
and refrigerated at 4◦ C until they were analyzed. Sampling
90 standard methods were used for each parameter.[16]
Physical and chemical parameters
We measured BOD5 , COD, conductivity, DO, fats and oils
(FO), MBAS, pH, phenols, total nitrogen, temperature (T),
TDS, TSS, and total solids (TS) in both effluents and in-
95 fluents of the seven wastewater treatment plants investi-
gated during the 2006 dry and rainy season using standard
methods.[16]
Metal determination
To determine the concentration of the metals (Al, As,
100 Cd, Cr, Cu, Fe, Pb, Mn, Hg and Zn) in the samples
we used atomic absorption (AA), according to standard
methods.[16] Air-acetylene flame (method 3111B) was used
to determine Cr, Cu, Fe, Mn and Zn. Nitrous oxide-
acetylene flame (3111D) was used for Al. In order to
105 increase the sensibility of both flame methods, samples
were concentrated 10-fold during the digestion procedure
(3030E). The detection limits for Al, Cr, Cu, Fe, Mn and
Zn were 130, 14, 23, 25, 12 and 5µg/L respectively. An
electrothermal method (3113 B) was used to determine Cd
110 and Pb (detection limits of 0.05 and 1.5 µg/L respectively).
Hydride generation (3114) and cold-vapor (3112) methods
were used to make As and Hg determinations (detection
limits of 0.1 and 0.3 µg/L, respectively). A Perkin-Elmer
AAnalyst 100 model AA equipment was used.
115 Quality assurance
A standard curve correlation coefficient in each heavy met-
als group of analysis was used as a quality control. The
standard curve correlation coefficient was higher than 0.98.
Internal standard and spiked samples recovery percentages
120 were between 80 and 110%. Duplicated samples coefficient
of variation was always smaller than 15%. J. T. Baker stan-
dards were used for atomic absorption. The analyses of all
the standard AA solutions were verified against identifiable
U.S. National Institute for Standardization and Technol-
125 ogy (NIST) solutions. Each sample was analyzed in tripli-
cate.
Daphnia magna 48-h acute toxicity test
We used the D. magna acute toxicity protocol detailed in the
Mexican Norm.[6] Briefly, this technique consisted of 48-h
3
exposure of 24-h old neonates of D. magna to a control 130
and 6 different concentrations determined through a range
toxicity test.[17] In the control, 10 neonates were placed
in each of 3 vials with 100 mL of EPA medium.[18] The
same was done for each treatment except that, besides the
EPA medium, the vials contained the corresponding test 135
concentration (typically 100%, 50%, 25%, 12.5%, 6.25%
dilutions of the wastewater). No food was given to the
neonates during this 48-h period in an incubator with a
18:6 L:D schedule. Light intensity was kept between 400
to 1000 lux as determined by an illuminometer (Kyoritan 140
Electrical Instruments), and temperature was kept at 20
± 1◦ C as determined by a microcomputer thermometer
(Hanna Instruments). Lack of movement was the endpoint
used to score the mortality of the animals.
48-h acute toxicity tests using Lecane quadridentata 145
We used Lecane quadridentata collected from Lake
Chapala, Mexico.[9] Lecane quadridentata was continuously
cultured in EPA medium[19] and fed Nanochloris oculata
(UTEX strain LB2194). Asexual eggs were collected and
incubated at 25◦ C in Petri dishes with EPA medium. EPA 150
medium had pH 7.4–7.8 and it hardness was 80–100 mg/L
CaCO3 . Acute toxicity tests were conducted in 24-well
polystyrene plates (Costar Co, U.S.A.), following the proto-
col of Pérez-Legaspi and Rico-Martı́nez.[9] Briefly, 10 24-h
old neonates were placed in each well containing 1.0 mL 155
EPA medium and incubated for 48-h at 25◦ C in darkness.
Besides a negative control, we used copper as our positive
control, and up to 7 wastewater dilutions. Each experi-
ment included 3 replicates. The test was concluded after
48-h, at which time the number of immobilized rotifers was 160
recorded and the LC50 was determined.
Statistical analysis for the 48-h acute toxicity test
Data were analyzed by means of the software DL50 to ob-
tain LC50 probit values. Statistica 6.0 (Stat-Soft Inc., 1993)
was used to determine r2 values. Tests were considered valid 165
only if mortality was less than 10% in the control.
Calculations of acute toxicity units
Median lethal concentration (LC50) units were trans-
formed into acute Toxicity Units (aTU) according with
Movahedian et al.[20] and Sánchez-Meza et al.[5] using Equa-
tion 1.
aTU = 100/LC50% (v/v) (1)
To calculate the toxicity removal efficiency Equation 2
was employed:
E = (aTUi − aTUe)/aTUi × 100 (2)
Where: E = Efficiency (%), aTUi = Acute toxicity units
of the influent, aTUe = Acute toxicity units of the effluent.
 4 Guzmán et al.

170 The toxicity level of each effluent and influent was de- Lecane quadridentata 48-h acute toxicity tests
termined according to the aTU obtained from the acute
In the case of test toxicity assessed with Lecane quadriden-
toxicity tests with both D. magna and L. quadridentata,
tata, CL50 aTU fluctuated between 2.6 and 8.3 in influents
and using the classification proposed by CETESB.[20,21]
during the dry season, a range that is considered as toxic 220
or very toxic (Table 6). In the rainy season the CL50 aTU
ranged from 3.9 to 8.3 being toxic to very toxic. In fact,
Results this latest value was the highest toxicity value found in this
study (Table 7). In contrast, the effluents in the dry season
175 Physical and chemical parameters fluctuated from 1.6 to 2.1, being moderately toxic to toxic. 225
In the rainy season the CL50 aTU ranged between 1.6 and
The monitoring results regarding physical and chemical
2.6 being moderately toxic to toxic (Table 6). Typical regres-
parameters of both influents and effluents during the dry
sion graphs of LC50 values obtained from environmental
season are shown in Table 1. On the 2 lower rows in Table 1
samples for L. quadridentata are shown in Figures 2b-e.
are shown the maximum allowed limits (MAL) established
The r2 values for this species were between 0.90 and 0.99. 230
180 by Mexican guidelines for each parameter. The conductiv-
ity, DO, pH and temperature of the influents were within
the MALs. The influents had values above the MALs for Toxicity removal efficiency
MBAS and total nitrogen. The COD, conductivity, DO,
The toxicity removal efficiency or the percentage of aTU
fats and oil, MBAS, pH, temperature and TSS of the ef-
removed from each wastewater treatment plant, was cal-
185 fluents were within the MALs. The BOD5 , total nitrogen,
culated applying equation 2. Obtained values are shown
TDS and TSS were above the MALs in some effluents dur-
in Table 7. In the dry season the%aTU removal for the 7 235
ing the dry season. In Table 2, the results of the parameters
wastewater treatment plants fluctuated between 59.1% and
during the rainy season are shown. The BOD5 , MBAS,
98.0% when Daphnia magna was employed. The removal
and total nitrogen values were above the MALs in almost
toxicity value using Lecane quadridentata showed a range
190 all influents. Only a few TDS and TSS values were above
from 22.1% to 74.6%. In the rainy season the values ranged
the MALs in influents. Only few TDS and total nitrogen
from 80.2% to 93.9% for Daphnia magna, and from 49.3% 240
values were above the MALs in effluents.
to 75.6% for Lecane quadridentata.

Metal analysis
Discussion
The metal concentration results are shown in Tables 3 and
195 4. In the dry season the values for Al, Fe, Zn, Mn, Cr, Hg In general, the most important wastewater treatment plants
and Pb in influents were above the MALs (Table 3). The in the state of Aguascalientes efficiently removed toxicity.
values of Fe, Mn and Pb in effluents were above the MALs However, this first statement has to be dissected to under- 245
in the dry season (Table 3). In the rainy season the Mn, Cr stand its implication. First, this result is especially signifi-
and Hg values in influents were above the MALs. Only Mn cant since wastewater treatment plants in Mexico were not
200 concentration was above the MALs in effluents (Table 4). particularly designed to remove toxicity. In fact, legislation
about toxicity in Mexico is somewhat unclear. Most of the
guidelines that apply to wastewater treatment plants focus 250
Daphnia magna 48-h acute toxicity tests on biological (fecal bacteria), chemical (BOD5 , COD) or
physical (conductivity, pH, temperature) parameters. Re-
The results of the 48-h acute toxicity tests with Daphnia
moval of biological, chemical, and physical parameters is
magna are shown in Table 5. The LC50 values are shown as
included in Mexican official guidelines.
aTU. In the D. magna tests the CL50 aTU of the influents
The pH is a valuable environmental variable, yet it is dif- 255
205 ranged from 0.3 to 4.5. That is, from slightly toxic to very
ficult to assess its impact on a particular reservoir since pH
toxic[21,22] in the dry season. On the other hand, the influ-
is influenced by many factors.[29] The pH influences toxic-
ents were more homogeneous in toxicity during the rainy
ity and availability of many compounds.[30] All pH values
season. The CL50 aTU ranged from 1.9 to 2.8 and were
recorded in this work fit with the Water Quality Mexican
classified from moderately toxic to toxic (Table 5). Regard-
Ecological Criteria[27] and with the values recommended 260
210 ing the effluents in the dry season, the LC50 aTU varied
by Wenner and Geist[31] (Tables 1 and 2). The relationship
from 0.1 to 0.4 and were considered slightly toxic. In the
among conductivity, total dissolved solids (TDS) and the
rainy season the LC50 aTU of the effluents ranged from 0.1
concentrations of several metallic ions and the contribution
to 0.4 and were also classified as slightly toxic (Table 5). A
of these 3 parameters to general toxicity have influenced the
typical regression graph of LC50 values obtained with en-
U.S. Environmental Protection Agency (EPA) rule on con- 265
215 vironmental samples for Daphnia magna is shown in Figure
trolling conductivity to a maximum 3000 µS/cm.[12] Three
2a.
 Table 1. Chemical and physical parameters of the 7 wastewater treatment plants investigated; data correspond to the 2006 dry season.

Influents

Temp. Cond. D.O. BOD5 COD Total N TTS TSS TDS MBAS Phenols F and O
Plant (◦ C) pH (µScm−1 ) (mgL−1 ) (mgL−1 ) (mgL−1 ) (mgL−1 ) (mgL−1 ) (mgL−1 ) (mgL−1 ) (mgL−1 ) (mgL−1 ) (mgL−1 )

RR 27.00 7.55 1110.00 0.25 234.00* 1111.00* 240.00* 1295.00* 495.83* 799.17 19.12 0.248 12.24
PA 27.00 7.82 1178.00 0.12 119.00* 694.50 140.00* 815.00 240.00* 575.00 92.82* 0.174 7.35
SFR 26.00 7.27 1799.00 0.23 90.00* 771.00 156.00* 780.00 220.00* 560.00 84.83* 0.176 7.45
JM 26.00 7.38 3800.00 0.36 98.00* 608.00 135.00* 1070.00 140.00 930.00* 80.31* 0.177 6.42
LS 26.00 7.15 6960.00* 0.78 149.00* 941.00 110.00* 945.00 120.00 825.00 98.73* 0.172 9.41
AGS 26.00 7.26 9450.00* 0.13 254.03* 557.50 160.00* 1140.00 140.00 1000.00* 26.42* 0.334 0.74
PIVA 26.00 7.37 1540.00 0.12 211.00* 1392.50* 260.00* 2025.00* 180.00 1845.00* 112.99* 0.161 71.57*
Effluents
RR 26.00 7.69 1300.00 4.22 20.00 23.10 3.00 535.00 36.00 499.00 0.19 0.032 0.07
PA 27.00 6.99 951.00 2.09 20.00 14.30 8.00 525.00 123.00 402.00 0.17 0.033 0.08
SFR 26.00 7.37 1464.00 3.57 20.00 21.50 8.00 645.00 51.00 594.00 0.14 0.032 1.54
JM 25.00 7.48 3900.00 3.34 20.00 31.40 16.00 920.00 34.00 886.00* 0.40 0.034 0.06
LS 25.00 7.63 679.00 4.41 20.00 34.00 14.00 635.00 21.00 614.00 0.43 0.034 0.29
AGS 27.00 7.39 817.00 3.67 100.00* 159.30 101.00* 900.00 80.00 820.00 0.96 0.043 0.44
PIVA 27.00 7.94 1323.00 4.80 20.00 117.50 27.00 1290.00* 18.00 1272.00* 1.44 0.037 1.39
MAL (40.00) (6.5–8.5) (5000.00) (5.00) (30.00) (1000.00) (40.00) (1200.00) (200.00) (850.00) (20.00) (3.50) (15.00)
[35] [28] [37] [28] [35] [36] [35] [36] [35] [36] [36] [38] [35]
Authority

NOM-001-ECOL[35] ; CE-CCA-001/89[28] ; Tchobanoglous and Burton[36] ; NOM-CCA-031-ECOL/1993[37] ; USEPA, 2002[38] .

*Values above the MALs.
Temp. = Temperature, Cond. = Conductivity, D. O. = Dissolved Oxygen, BOD5 = Biochemical Oxygen Demand, COD = Chemical Oxygen Demand, Total N = Total nitrogen, TTS =
Total Total Solids, TSS = Total Suspended Solids, TDS = Total Dissolved Solids, MBAS = Methylene Blue Active Substances, F and O = Fats and Oils, MAL = Maximum Allowed
Levels, ND = No Detected.
Abbreviations of the wastewater treatment plants correspond to: RR = Rincón de Romos, PA = Pabellón de Arteaga, SFR = San Francisco de los Romo, JM = Jesús Marı́a, LS = Los
Sauces, AGS = Ciudad de Aguascalientes, PIVA = Parque Industrial del Valle de Aguascalientes.

5
6
Table 2. Chemical and physical parameters of the 7 wastewater treatment plants investigated; data correspond to the 2006 rainy season.

Influents

Temp. Cond. D.O. BOD5 COD Total N TTS TSS TDS MBAS Phenols F and O
Plant (◦ C) pH (µScm−1 ) (mgL−1 ) (mgL−1 ) (mgL−1 ) (mgL−1 ) (mgL−1 ) (mgL−1 ) (mgL−1 ) (mgL−1 ) (mgL−1 ) (mgL−1 )

RR 28.00 7.48 1245.00 0.28 241.94* 714.00 90.00* 1650.00* 295.00* 1355.00* 19.12 0.120 12.24
PA 26.00 7.61 1042.00 0.72 302.42* 916.00 62.00* 1055.00 430.00* 625.00 92.82* 0.144 7.35
SFR 26.00 7.44 1103.00 0.39 535.35* 2088.00* 167.00* 1650.00* 225.00 1425.00* 84.83* 0.152 7.45
JM 25.40 7.68 1261.00 0.31 185.30* 426.00 157.00* 995.00 300.00* 695.00 80.31* 0.150 6.42
LS 26.60 8.04 1235.00 0.10 302.42* 746.00 190.00* 1190.00 195.00 995.00* 98.73* 0.147 9.41
AGS 25.00 7.29 1525.00 0.31 244.03* 770.00 160.00* 1115.00 300.00* 815.00 26.42* 0.125 0.74
PIVA 26.00 7.46 1078.0 0.21 210.00* 1393.00* 240.00* 2025.00* 180.00 1845.00* 112.99* 0.149 71.57*
Effluents
RR 25.00 7.67 841.00 3.75 ND 22.00 49.00* 670.00 90.00 580.00 0.19 0.050 0.07
PA 23.00 7.52 839.00 1.64 ND 3.00 43.00* 725.00 30.00 695.00 0.17 0.040 0.08
SFR 24.00 7.54 875.00 1.66 67.11* 85.00 41.00* 435.00 147.50 287.50 0.14 0.050 1.54
JM 24.00 7.68 1320.00 2.70 ND 29.00 96.00* 1055.00 50.00 1005.00* 0.40 0.033 0.06
LS 25.50 7.06 979.00 2.52 ND 17.00 62.00* 835.00 97.50 737.50 0.43 0.048 0.29
AGS 25.00 7.74 1282.00 3.43 100.70* 165.00 101.00* 885.00 30.00 855.00* 0.96 0.058 0.44
PIVA 26.00 7.47 962.00 3.59 18.00 118.00 56.00* 1290.00* 18.00 1272.00* 1.44 0.051 1.39
MAL (40.00) (6.5–8.5) (5000.00) (5.00) (30.00) (1000.00) (40.00) (1200.00) (200.00) (850.00) (20.00) (3.50) (15.00)
[35] [28] [37] [28] [35] [36] [35] [36] [35] [36] [36] [38] [35]
Authority

NOM-001-ECOL[35] ; CE-CCA-001/89[28] ; Tchobanoglous and Burton[36] ; NOM-CCA-031-ECOL/1993[37] ; USEPA, 2002[38] .

*Values above the MALs.
Temp. = Temperature, Cond. = Conductivity, D.O = Dissolved Oxygen, BOD5 = Biochemical Oxygen Demand, COD = Chemical Oxygen Demand, Total N = Total nitrogen, TTS =
Total Total Solids, TSS = Total Suspended Solids, TDS = Total Dissolved Solids, MBAS = Methylene Blue Active Substances, F and O = Fats and Oils, MAL = Maximum Allowed
Levels, ND = No Detected.
Abbreviations of the wastewater treatment plants correspond to: RR = Rincón de Romos, PA = Pabellón de Arteaga, SFR = San Francisco de los Romo, JM = Jesús Marı́a, LS = Los
Sauces, AGS = Ciudad de Aguascalientes, PIVA = Parque Industrial del Valle de Aguascalientes.
Wastewater in seven treatment plants in Aguascalientes, Mexico 7
Table 3. Metal concentrations of the 7 wastewater treatment plants investigated. Data correspond to the 2006 dry season.

Influents
Plant Al Fe Zn Mn Cr Cu Cd As Hg Pb

RR 3.24 3.61 0.75 0.89* 0.05 0.13 ND 0.01 ND 0.17

PA 3.73 2.30 0.32 0.13 0.02 0.06 ND 0.02 ND 0.19
SFR 4.54 4.70 0.84 0.36* 0.03 0.18 ND 0.02 ND 0.16
JM 7.65* 49.20* 2.95* 1.69* 0.24 0.44 ND 0.01 ND 0.22*
LS 8.03* 5.45* 0.92 0.36* 0.05 0.33 ND 0.01 ND 0.17
AGS 4.30 2.01 1.04 0.42* 0.19 0.39 ND ND ND 0.25*
PIVA 5.52* 7.24* 2.42* 0.72* 1.28* 0.26 ND ND 0.11* 0.22*
Effluents
RR 0.04 0.18 0.13 0.02 0.02 ND ND ND ND 0.14
PA 0.08 0.26 0.20 ND 0.01 ND ND 0.01 ND 0.07
SFR 4.48 4.79 0.67 0.18 0.08 0.09 ND 0.01 ND 0.17
JM 0.12 0.51 0.37 0.01 0.06 0.04 ND 0.01 ND 0.22*
LS 0.22 0.46 0.66 0.03 0.02 0.06 ND 0.01 ND 0.22*
AGS 1.05 1.80 0.32 0.42* 0.06 0.05 ND 0.01 ND 0.17
PIVA 0.38 5.09* 0.39 0.31* 0.09 0.08 ND 0.01 ND 0.12
MAL 5.00 5.00 2.00 0.20 1.00 1.00 0.20 1.00 0.02 0.20
[28] [28] [28] [36] [35] [28] [35] [28] [35] [35]
Authority

NOM-001-ECOL[35] ; CE-CCA-001/89[28] ; Tchobanoglous and Burton[36] .

*Values above the Maximum Allowed Limits (MAL).
ND = No Detected.
Abbreviations of the wastewater treatment plants correspond to: RR = Rincón de Romos, PA = Pabellón de Arteaga, SFR = San Francisco de los
Romo, JM = Jesús Marı́a, LS = Los Sauces, AGS = Ciudad de Aguascalientes, PIVA = Parque Industrial del Valle de Aguascalientes. All values
are in mgL−1 .

Table 4. Metal concentrations of the seven wastewater treatment plants investigated. Data correspond to the 2006 rainy season.

Influents
Plant Al Fe Zn Mn Cr Cu Cd As Hg Pb

RR 0.34 0.35 0.07 1.23* ND ND 0.02 0.16 ND ND

PA 1.40 1.26 0.14 0.30* ND ND 0.01 0.23 ND ND
SFR 3.01 7.80 0.31 0.33* ND ND ND 0.20 ND ND
JM 0.53 0.49 0.08 0.41* ND ND 0.01 0.12 ND ND
LS 0.39 0.43 0.14 0.35* ND ND ND 0.10 ND ND
AGS 0.31 1.55 0.06 0.42* 0.06 0.26 ND ND 0.11* ND
PIVA 0.92 0.63 0.53 0.87* 1.90* ND 0.13 ND ND ND
Effluents
RR ND 0.05 ND 0.04 ND ND 0.01 0.17 ND ND
PA ND 0.06 ND 0.04 ND ND ND 0.35 ND ND
SFR 0.28 0.47 0.05 0.02 ND ND 0.01 0.18 ND ND
JM ND 0.10 0.03 0.03 ND ND ND 0.10 ND ND
LS ND 0.06 0.03 0.24* ND ND ND 0.05 ND ND
AGS ND 0.12 0.01 0.31* ND 0.05 ND ND ND ND
PIVA ND 0.41 ND ND ND ND ND 0.08 ND ND
MAL 5.00 5.00 2.00 0.20 1.00 1.00 0.20 1.00 0.02 0.20
[28] [28] [28] [36] [35] [28] [35] [28] [35] [35]
Authority

NOM-001-ECOL[35] ; CE-CCA-001/89[28] ; Tchobanoglous and Burton[36] .

*Values above the Maximum Allowed Limits (MAL).
ND = No Detected.
Abbreviations of the wastewater treatment plants correspond to: RR = Rincón de Romos, PA = Pabellón de Arteaga, SFR = San Francisco de los
Romo, JM = Jesús Marı́a, LS = Los Sauces, AGS = Ciudad de Aguascalientes, PIVA = Parque Industrial del Valle de Aguascalientes. All values
are in mgL−1 .
 8 Guzmán et al.
Table 5. Acute toxicity tests (48-h) with Daphnia magna; data Table 6. Acute toxicity tests (48-h) with Lecane quadridentata;
correspond to the 2006 dry and rainy seasons. data correspond to the 2006 dry and rainy seasons.

Influents Influents
Dry season Rainy season Dry season Rainy season
Water Water
treatment LC50 Level of LC50 Level of treatment LC50 Level of LC50 Level of
plant LC50 (aTU) toxicity LC50 (aTU) toxicity plant LC50 (aTU) toxicity LC50 (aTU) toxicity

RR 35.7 2.8 T 49.5 2.0 T RR 20.6 4.9 VT 25.4 3.9 T

PA 151.3 0.7 ST 45.3 2.2 T PA 26.7 3.7 T 17.8 5.6 VT
SFR 57.2 1.8 MT 49.5 2.0 T SFR 23.7 4.2 VT 13.2 7.6 VT
JM 228.1 0.4 ST 49.5 2.0 T JM 38.2 2.6 T 23.9 4.2 VT
LS 400.0 0.3 ST 36.1 2.8 T LS 49.0 2.0 T 12.0 8.3 VT
AGS 61.0 1.6 MT 53.3 1.9 MT AGS 26.0 3.8 T 17.6 5.7 VT
PIVA 22.4 4.5 VT 51.2 2.0 T PIVA 12.0 8.3 VT 13.3 7.5 VT
Effluents Effluents
RR 549.8 0.2 ST 588.1 0.2 ST RR 60.2 1.7 MT 61.6 1.6 MT
PA 588.1 0.2 ST 306.3 0.3 ST PA 60.2 1.7 MT 47.3 2.1 T
SFR 286.9 0.4 ST 249.4 0.4 ST SFR 48.3 2.1 T 53.6 1.9 MT
JM 549.2 0.2 ST 750.8 0.1 ST JM 61.7 1.6 MT 47.0 2.1 T
LS 1151.1 0.1 ST 588.1 0.2 ST LS 38.2 2.6 T 44.8 2.2 T
AGS 400.0 0.3 ST 400.4 0.2 ST AGS 48.9 2.0 T 37.8 2.6 T
PIVA 1151.1 0.1 ST 588.1 0.2 ST PIVA 47.3 2.1 T 47.2 2.1 T

ST = Slightly Toxic, MT = Moderately Toxic. T = Toxic and VT =
Very Toxic. Classification taken from Brazil’s Compañı́a de tecnologı́a
de saneamento ambiental (CETESB), 1991[21] .
Abbreviations correspond to: RR = Rincón de Romos, PA = Pabellón
de Arteaga, SFR = San Francisco de los Romo, JM = Jesús Marı́a,
LS = Los Sauces, AGS = Ciudad de Aguascalientes, PIVA = Parque
Industrial del Valle de Aguascalientes.
ST = Slightly Toxic, MT = Moderately Toxic. T = Toxic and VT =
Very Toxic. Classification taken from Brazil’s Compañı́a de tecnologı́a
de saneamento ambiental (CETESB).[21]
Abbreviations correspond to: RR = Rincón de Romos, PA = Pabellón
de Arteaga, SFR = San Francisco de los Romo, JM = Jesús Marı́a,
LS = Los Sauces, AGS = Ciudad de Aguascalientes, PIVA = Parque
Industrial del Valle de Aguascalientes.

dry season influents showed the highest values of conduc-
tivity: JM, LS and AGS (3800, 6960 and 9450 µS/cm,
respectively). These high conductivity levels were matched
270 with high values of TDS (Tables 1 and 2). This is relevant,
because according to the U.S. EPA[12] , if conductivity ex-
ceeds 1000 µS/cm, TDS can influence the toxicity param-
eters of an effluent. Our results showed that all influents
and four effluents exceeded this 1000 µS/cm value in dry
275 season. These conductivity values are explained in part by
high concentrations of metallic ions.
The DO has great importance for aquatic organisms.
Whitledge[27] defined a reservoir as hypoxic when DO con-
centration is less than 2.0 mgL−1 , and anoxic when DO is
280 0.0 mgL−1 . The Mexican ecological criteria[28] established
5.0 mgL−1 as a recommended value for the protection of
aquatic life. In the present work the mean DO value for in-
fluents was 0.28 mgL−1 and for effluents the value was 3.73
mgL−1 (both in the dry season), which were still below the
285 recommended value (Tables 1 and 2).
Regarding the chemical and physical parameters in the
dry season, the wastewater entering the treatment plants
had a BOD5 concentration that exceeded the MAL (30 mg
L−1 ) 3.0 to 8.5 times. After the plants treated wastewater,
290 BOD5 values were still above the MAL in only one plant
(AGS). In the rainy season, the influents had a BOD5 con-
centration that exceeded MAL 6.1 to 17.8 times. Similarly
BOD5 concentration in treated wastewater was 2- to 3- fold
higher with respect to MAL in two wastewater plants (SFR
and AGS) (Tables 1 and 2). 295
Total nitrogen exceeded the MAL (40 mgL−1 ) in the dry
season influents and the recorded values were from 2.8-
to 6.5-fold higher than the MAL. The dry season effluent
of the city of Aguascalientes wastewater treatment plant
(AGS) exceeded by 2.5-fold the total nitrogen MAL (Tables 300
1 and 2). The contribution of treated wastewater enriched
with organic matter and nitrogen can contribute to the
eutrophication of the San Pedro River.
With the inclusion of the metal data in this article, it is
important to point out the potential causes of the toxicity 305
found in the samples of both treated and untreated water.
Al, Fe, Zn and Pb were found in several environmental
samples at concentrations that exceed the MAL and these
concentrations exceed the LC50 values for each particu-
lar metal obtained for L. quadridentata (Rico-Martı́nez, 310
2009, unpublished data). In the dry season PIVA influent
exceeded the Al, Fe, Zn, Mn, Cr, Hg and Pb MAL values.
In JM influent Al, Fe, Zn, Mn and Pb were above the MAL.
Mn and Pb exceeded the MAL values in AGS wastewater
treatment plant influents. The LS wastewater plant had Al, 315
Fe and Mn values above the MAL values in influents. SFR
Wastewater in seven treatment plants in Aguascalientes, Mexico 9

Y = -15.338 + 0.0081X; r2 = 0.7946

(a) LC50 = 61.002
120

100

% Mortality 80

60

40

20

-20
-20 0 20 40 60 80 100 120
% Dilution

Y = 0.3857 + 1.9161X; r2 = 0.9743

(b) LC50 = 26.035
120

100

80
% Mortality

60

40

20

-20
-10 0 10 20 30 40 50 60
% Dilution

2
Y = 1.1383 + O.9969X; r = 0.9964
(c)
LC50 = 48.954
120

100

80
% Mortality

60

40

20

-20
-20 0 20 40 60 80 100 120
% Dilution

Fig. 2. CL50 regression graphs. (a) A typical D. magna CL50 regression graph from the sample collected at the City of Aguascalientes
WTP influent. (b) L. quadridentata CL50 regression graph collected from the City of Aguascalientes WTP influent. (c) L. quadridentata
CL50 regression graph collected from the City of Aguascalientes WTP effluent. (d) L. quadridentata CL50 regression graph collected
from the PIVA WTP influent. (e) L. quadridentata CL50 regression graph collected from the PIVA WTP effluent.
 10 Guzmán et al.

(d) Y = 5.4973 + 3.6801X; r2 = 0.9796

LC50 = 12.037
120

100

80

60
% Mortality

40

20

-20
-2 0 2 4 6 8 10 12 14 16 18 20 22 24 26
% Dilution

Y = 0.5802 + 1.0533X; r 2 = 0.9637

(e)
LC = 47.298
120

100

80

60
% Mortality

40

20

-20
-20 0 20 40 60 80 100 120
% Dilution

Fig. 3. (continued)

and RR had Mn concentrations above the MAL in influ-
ents. All influents exceeded Mn MAL in the rainy season.
The AGS influent had Hg values above the MAL value,
320 and the PIVA influent had Mn, Cr concentrations above
the MAL in the rainy season (Tables 3 and 4). These results
on metal determination correspond well with a previous
study of metals along the San Pedro River.[32]
When D. magna was used as a model organism, toxi-
325 city units for influents ranged from 0.3 to 4.5 and from
0.1 to 0.4 for effluents (Tables 5). These values for efflu-
ents are acceptable according to international guidelines,
and they compare well with data from Villegas-Navarro et
al.[26] who reported values for D. magna of 1.5 to 7.2 aTU
330 for treated textile wastewater from Puebla, Mexico. How-
ever, when L. quadridentata was the model organism, they
ranged from 2.0 to 8.3 in influents and from 1.6 to 2.6 in ef-
fluents (Table 6). This treated water that discharges into the
San Pedro River would be considered as “toxic” according
335 to CETESB.[21]
The dry season in Aguascalientes includes the months
from November to February and the rainy season includes
the months from August to October. As expected, the influ-
ents showed more toxicity than the effluents, and there was
more toxicity in the rainy than in the dry season. These re- 340
sults are in agreement with the findings of Santos-Medrano
et al. [15] The authors used samples collected along the San
Pedro River and its main tributaries for 2003–2004.
Mexican legislation includes 48-h Daphnia magna acute
tests as a standardized probe. Being a technical norm, it is 345
a step down from mandatory official norms in the Mexi-
can legislation. In our results with D. magna the removal
efficiency of the wastewater treatment plants had average
values of 79.1% in the dry season and 88.7% in the rainy
season. Furthermore, the plant that is located in the city 350
of Aguascalientes, treats more wastewater that all other six
plants together, and had efficiencies of 84.7% and 86.6%
in dry and rainy season, respectively (Table 7). Therefore,
the toxicity removal with D. magna seems to be efficient
 Wastewater in seven treatment plants in Aguascalientes, Mexico 11
Table 7. Differences in toxicity removal among the 7 wastewater treatment plants investigated, using the 48-h acute tests with either
Daphnia magna or Lecane quadridentata; data correspond to the 2006 dry and rainy seasons.

Dry season Dry season Differences in removal Rainy season Rainy season Differences in removal
Treatment D. magna L. quadridentata Dry season D. magna L. quadridentata Rainy season
plant aTU (%) aTU (%) aTU (%) aTU (%) aTU (%) aTU (%)

RR 93.6 65.8 27.8 91.6 58.8 32.8

PA 74.2 55.6 18.6 85.0 62.5 22.5
SFR 80.0 50.8 29.2 80.2 75.6 4.6
JM 59.1 38.2 20.9 92.1 49.3 42.8
LS 64.0 22.1 41.9 93.9 73.2 20.7
AGS 84.7 46.9 37.8 86.6 53.5 33.1
PIVA 98.0 74.6 23.4 91.3 71.9 19.3
Mean
Abbreviations correspond to: RR = Rincón de Romos, PA = Pabellón de Arteaga, SFR = San Francisco de los Romo, JM = Jesús Marı́a, LS =
Los Sauces, AGS = Ciudad de Aguascalientes, PIVA = Parque Industrial del Valle de Aguascalientes.

355 and conforms, in general, to the correspondent Mexican
Norms (see also chemical and physical parameters, Tables
1 and 2). The toxicity removal efficiencies obtained with
L. quadridentata were 47.7% and 63.5% in dry and rainy
season respectively.
360 The observed difference in sensitivity between D. magna
and L. quadridentata is especially significant if we con-
sider the fact that D. magna has never been found as a
natural inhabitant of Mexican reservoirs[23] , and that L.
quadridentata has shown to be far more sensitive than D.
365 magna in several experiments and environmental toxicity
surveys.[15,24,25] These high/intermediate toxicity removal
efficiencies should contribute over the next few years to
clean the San Pedro River, an area that until now[15] has
been heavily contaminated. The PIVA wastewater treat-
370 ment plant showed the highest toxicity removal efficiencies
with values above 90.0% for dry and rainy seasons when
D. magna was used. Although this plant only treats 28.78
Ls−1 , it had the most toxic discharge in terms of CL50 aTU
to the San Pedro River.[15]
375 CETESB[21] classifies effluents toxicity in four levels:
slightly toxic, moderately toxic, toxic and very toxic, ac-
cording with their aTU (<1.33, 1.33–1.99, 2.00–4.00 and
>4.00, respectively). When we compare the a TU of the all
28 influent and effluent samples corresponding to dry and
380 rainy seasons, using D. magna and L. quadridentata, there
was a clear difference in the levels of toxicity for a partic-
ular influent or effluent. In 89.3% of the cases, L. quadri-
dentata was more sensitive than D. magna. Furthermore,
11 of 28 of these samples (39.3%) ranged from “moderately
385 toxic” to “very toxic” for D. magna, but 100.0% of the sam-
ples ranged from “moderately toxic” to “very toxic” for L.
quadridentata. If we compare the toxicity level of D. magna
with the corresponding toxicity level of L. quadridentata,
50.0% of total samples changed their toxicity two levels
390 higher. For example, the toxicity level for the SFR wastew-
ater treatment plant influent was “moderately toxic” in dry
season when it was tested with D. magna, and the toxicity
level for the same influent with L. quadridentata was “very
toxic”. Therefore, the toxicity level was modified two lev-
els higher (Tables 5 and 6). Rico-Martı́nez[33] has already 395
reported a higher sensitivity of L. quadridentata compared
with D. magna.
According to Baudouin and Scoppa[34] , the choice of
zooplanktonic organisms as models for toxicity tests (when
treated wastewater discharged into reservoirs is assessed), 400
is of great utility to establish ecological criteria when the
ultimate goal is the preservation of aquatic ecosystems.
D. magna and L. quadridentata are important zooplank-
tonic organisms essential components of freshwater webs.
The environmental stress produced on these populations 405
could lead to important damages to the entire web, or
to the complete aquatic ecosystem. When we compared
the toxicity level among the effluents discharging to the
San Pedro River between both model organisms, we found
that with D. magna all effluents were classified as “slightly 410
toxic”. However, when the same effluents were tested with
L. quadridentata, they were classified as “moderately toxic”
or “toxic”. This implies two things: first, that L. quadri-
dentata is more sensitive than D. magna as a toxicity
test model organism for surveys, and second that toxic 415
effluents are being discharged to the San Pedro River
from the wastewater treatment plants (including impor-
tant loads of total nitrogen). Then, the privileged use of D.
magna by the Mexican Legislation can produce underes-
timation of actual toxicity leading to erroneous decision- 420
making that might jeopardize the future of the San Pedro
River.
Conclusion
The seven wastewater treatment plants investigated effi-
ciently removed toxicity (79.1% in the dry season and 88.7% 425
in the rainy season) when Daphnia magna was used as model
organism, but perform poorly when the freshwater rotifer
 12
Lecane quadridentata was employed (47.7% in dry season
and 63.5% in rainy season). Toxicity was greater during the
430 rainy season in all seven plants and this result is in agree-
ment with previous studies of toxicity along the San Pedro
River.
We found that the COD, conductivity, DO, pH, phenols,
MBAS, temperature, and TSS were within the MALs in the
435 effluents during the rainy season. Whereas the BOD5 , total
nitrogen and TDS showed levels greater than the MAL in
effluents during the dry season. The levels of BOD5 , MBAS,
and total nitrogen were greater than the MAL values in
rainy season in 6 of 7 studied plant influents. On the other
440 hand, the values of TDS and TSS in influents were above the
MAL during the rainy season in 4 influents of 7 wastewater
plants. Total nitrogen is of particular interest since it was
above the MAL value in both influents and effluents in the
rainy season and can produce eutrophication to reservoirs
445 along the San Pedro River.
In the dry season the levels of Al, Fe, Zn, Mn, Cr, Hg
and Pb in some influents were higher than MAL values
and Fe, Mn and Pb in effluents were above the MAL in
the same season. During the rainy season only the levels of
450 Mn Cr and Hg in influents and those of Mn in effluents
were above the MAL. The presence of these metals is in
agreement with previous reports of metals found in the San
Pedro River. The results of the toxicity tests demonstrated
that the freshwater rotifer Lecane quadridentata was more
455 sensitive than the cladoceran Daphnia magna when exposed
to the influents and effluents of the wastewater treatment
plants (in 89.3% of the samples).
Acknowledgments
Authors want to thank: Natural Resources and Environ-
460 mental Ministry (SEMARNAT) and Science and Technol-
ogy National Council (CONACyT) of Mexico for their
support. FOSEMARNAT-2004-01-78 grant. F. Torres-
Guzmán, gives thanks to CONACyT for the 198929 Schol-
arship, and to Martha Evelia Pérez Reyes, Laura Ya-
465 mamoto Flores, Jenniffer López Baltazar, Samanta Ramos
Gómez and Guilda Guzmán Colis, Jorge Hernández
Sánchez, who are members of the Biotechnology and
Environmental Studies Laboratories of the Universidad
Autónoma de Aguascalientes. The help received from Ma.
470 Guadalupe González-Maldonado a doctoral student of
Mexico’s Instituto Politécnico Nacional is especially ap-
preciated.
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