Aquaculture Research, 2011, 42, 1415^1423 doi:10.1111/j.1365-2109.2010.02730.

Bioremediation and reuse of shrimp aquaculture

effluents to farm whiteleg shrimp, Litopenaeus
vannamei : a first approach

Luis R Mart|¤ nez-Co¤rdova1, Jose¤ A Lo¤pez-El|¤ as1, Guadalupe Leyva-Miranda2, Luis Armenta-Ayo¤n2 &
Marcel Martinez-Porchas3
1
Departamento de Investigaciones Cient|¤ ¢cas y Tecnolo¤gicas de la Universidad de Sonora, Sonora, Me¤xico
2
Maestr|¤ a en Biociencias, Universidad de Sonora, Sonora, Me¤xico
3
Centro de Investigacio¤n en Alimentacio¤n y Desarrollo, Sonora, Me¤xico

Correspondence: M Mart|¤ nez-Porchas, Centro de Investigacio¤n en Alimentacio¤n y Desarrollo, Km. 0.6 Carretera a La Victoria, Hermo-
sillo, Sonora, Me¤xico. E-mail: marcel@ciad.mx

Abstract to the global production of aquatic organisms grew

from 3.9% in 1970 to more than 36% in 2006 (FAO
Shrimp aquaculture e¥uents were bioremediated in
2009). The Crustaceans are the group with the high-
a two-phase system (System A) using the black clam
est growth rate (almost 17% per year from 2000 to
Chione £uctifraga and the benthic microalgae Navicu-
2006) and penaeid shrimp are by far the most impor-
la sp., and then reused to farm whiteleg shrimp Lito-
tant in terms of volume and value of production (FAO
penaeus vannamei. In the experimental design,
2009).
Systems B and C had an identical structure as System
However, the explosive development of shrimp
A, but no clams or microalgae were added. System B
aquaculture has caused some serious problems, such
received the same shrimp e¥uents while System C
as competition for water and land (PaŁez-Osuna, Gra-
received only estuarine water. Shrimp raw e¥uents
cia, Flores-Verdugo, Lyle-Fritch, Alonso-Rodr|¤ guez.,
had a poor water quality. System A improved
Roque & Ruiz-FernaŁndez 2003), environmental im-
the water quality by decreasing the concentrations
pacts, including deforestation, eutrophication of re-
of total nitrogen, total ammonia nitrogen (TAN), ni-
ceiving ecosystems, modi¢cation of habitat for
trites, nitrates, phosphates, total suspended solids
terrestrial and aquatic animals, modi¢cation of land-
(TSS) and organic suspended solids (OSS). System B
scape and hydrological patterns (GonzaŁlez-Ocampo,
also decreased the concentration of TAN, TSS
Morales, CaŁceres-Mart|¤ nez, Aguirre, HernaŁndez-VaŁz-
and OSS via sedimentation, but the e¡ect was less
quez, Troyo-Dieguez & Ortega-Rubio 2006; Thomas,
pronounced than that observed in System A. Shrimp
Courties, El Helwe, Herbland & Lemonnier 2010), the
reared in the bioremediated e¥uents (System A) had
dependence of formulated shrimp feed from ¢sh meal
better production (3166 kg ha  1) and higher survival
as the main protein ingredient (Tacon 2002) and the
(89.2%) than those reared in e¥uents from Systems B
continuous presence of epizooties (SaŁnchez-Mart|¤ -
(2610 kg ha  1,75.1%) and C (2874 kg ha  1, 82.1%). It
nez, Aguirre-GuzmaŁn & Mej|¤ a-Ruiz 2007).
is concluded that the bioremediation system was
As an example of the potential impact of aquacul-
moderately e⁄cient and the bioremediated e¥uents
ture e¥uents, Tacon (2002) shows data regarding
were suitable to farm L. vannamei.
how much organic matter (OM), nitrogen (N) and
phosphorous (P) is discharged into the environment
Keywords: bioremediation, shrimp e¥uent, Chione
for each tonne of shrimp harvested, depending on
£uctifraga, Navicula sp.
the feed conversion ratio (FCR). In Mexico, shrimp
aquaculture operates with a mean FCR of about 1.8;
Introduction
considering the annual production and the data pro-
Aquaculture has experienced vigorous growth vided by Tacon (2002), it is calculated that 112 mil-
worldwide in the last two decades. Its contribution lion kg of OM,7.8 million kg of N and 2.5 million kg of

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Bioremediation and reuse of e¥uents L R Mart|¤ nez-Co¤rdova et al.
P are discharged into the receiving ecosystems each
year (Mart|¤ nez-Co¤rdova, Mart|¤ nez-Porchas & Corte¤s-
Jacinto 2009). These are huge amounts and it is abso-
lutely necessary to stop and if possible to reverse this
process if we wish to have a sustainable aquaculture.
Some strategies have been used or suggested to
minimize these impacts, including settling lagoons
(Mart|¤ nez-Co¤rdova & Enriquez-Ocana 2007), septic
tank treatments (Summerfelt & Penne 2007), low or
zero water exchange (Balasubramanian, Pillai & Ra-
vichandran 2005), recirculation systems (Timmons,
Ebeling, Wheaton, Summerfelt & Vinci 2002; Leza-
ma-Cervantes, Paniagua-Michel & Zamora-Castro
2010), the use of mangrove forests as nutrient sinks
(Rivera-Monroy, Torres, Bahamon, Newmark & Twil-
ley 1999), polyculture practices (Martinez-Cordova &
Martinez-Porchas 2006; Mart|¤ nez-Porchas, Mart|¤ nez-
Co¤rdova, Porchas-Cornejo & Lo¤pez-El|¤ as 2010) and
bioremediation (Paniagua-Michel & Garcia 2003).
Bioremediation is the use of individual or
combined organisms (animal, vegetal, bacteria, etc.)
to minimize the polluting charge of e¥uents from
aquaculture or any other activity, taking advantage
of the natural or modi¢ed abilities of those organisms
to reduce and/or transform waste products (Chavez-
Crooker & Oberque-Contreras 2010). Bioremediation
can be conducted in di¡erent forms: in situ, ex situ,
biostimulation, bioagmentation and others. Some
examples of successful bioremediation practices are
the use of plants (phytoremediation), macroalgae,
microalgae, ¢lter feeders, bio¢lters (polymer spheres
with immobilized microorganisms), bio¢lms^bio£ocs
(De Schryver, Crab, Deforidt, Boon & Verstraete 2008;
Kuhn, Boardman, Craig, Flick Jr & Mclean 2009) or
combined systems including two or more of these
practices. Although it has been demonstrated that
some bivalves and micro- or macroalgae are capable
of bioremediating e¥uents, many of these studies
have been focused on the bioremediation of ¢sh e¥u-
ents (Hussenot 2003; Zhou,Yang, Hu, Liu, Mao, Zhou,
Xu & Zhang 2006; Liu,Wang & Lin 2010).
The use of endemic species of bivalve and microal-
gae to bioremediate e¥uents would prevent the intro-
duction of exotic species, which may cause other
problems. It is important to study di¡erent combina-
tions of these species in order to achieve the greatest
e⁄ciency.
The black clam (Chione £uctifraga) inhabits estu-
aries and shallow coastal waters in the Gulf of Cali-
fornia. It tolerates high concentrations of OM in the
water column and can withstand a wide range of
temperatures and salinities (Mart|¤ nez-Co¤rdova
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Aquaculture Research, 2011, 42, 1415–1423
1988), conditions that are similar to those prevailing
in shrimp farm e¥uent; also, the species has com-
mercial importance in north-western Mexico,
mainly as an artisanal ¢shery, but it is beginning to
be farmed in the region (Tinoco-Orta & CaŁceres-Mar-
t|¤ nez 2003). In the case of microalgae, Navicula sp. is a
diatom that has been found in shrimp ponds and can
be a food source for shrimp; also, it is reported to have
the ability to act as a bioremediator of water (Pania-
gua-Michel & Garcia 2003).
Based on the above information, the study was
focused on evaluating an integrated system using
benthic microalgae (Navicula sp.) and clam (C. £uctifra-
ga) to bioremediate shrimp aquaculture e¥uents and
reuse the bioremediated e¥uents to farm whiteleg
shrimp Litopenaeus vannamei at a microcosm level.
Material and methods
Organisms
The shrimp were obtained from a commercial farm
(Maricultura del Pac|¤ ¢co S.A., Mazatlan, Me¤xico)
and maintained under the laboratory conditions of
dissolved oxygen (DO) 6.05 mg L  1, temperature
28 1C, total ammonia nitrogen (TAN) 0.01mg L  1,
s
fed ad libitum (35% crude protein; Purina Me¤xico ,
Hermosillo, Me¤xico) and a daily water exchange of
100%. These conditions were maintained until the
shrimp achieved an individual biomass of 5 g.
Adult black clams (C. £uctifraga) with an average
size of 3.0  0.4 cm were hand collected from the La
Cruz estuary (28148 05700 N, 111155 03000 O); organisms
of a lower size were discarded, because we aimed to
evaluate only the capacity of adult clam, which have
a greater ¢ltration capacity. The clam was main-
tained under the above laboratory conditions for 2
weeks. During this period, the organisms were fed
with the diatom Chaetoceros muelleri.
The microalga Navicula sp. was obtained from an
aquaculture laboratory at Centro de Estudios Super-
iores del Estado de Sonora (CESUES, Navojoa, Sonora,
Me¤xico). The microalgae were scaled up from 10 mL
to 200 L using an F/2 medium with a double concen-
tration of silicates. Thereafter, the experimental units
were inoculated with the microalgae 1 week before
beginning the trial.
Shrimp culture system
The e¥uents used for the study were obtained from a
semiintensive culture of white shrimp L. vannamei
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Aquaculture Research, 2011, 42, 1415^1423 Bioremediation and reuse of e¥uents L R Mart|¤ nez-Co¤rdova et al.

(25 shrimp m  2) reared in tanks with estuarine
water.
The shrimp were farmed in six rectangular, plastic
tanks with an area of 6 m2 and ¢lled with water
pumped from the estuary; each tank was provided
with 0.9 m3 of sediment to achieve a height of 5^
6 cm. The culture conditions were as follows: time of
culture 50 days, initial size 5.0  0.3 g, stocking den-
sity 25 org m  2, feeding supply and frequency twice
a day to satiation (in feeding trays), the feed used was
Camaronina with 35% of crude protein (Purina Me¤x-
s
ico ) and the daily water exchange was 20%.
Bioremediation system
One half of the e¥uent was sent to a bioremediation
system and the other half was sent to an identical
physical system but was not treated. Both types of
water were then used to cultivate white shrimp.
The bioremediation system (System A) consisted of
two phases. In Phase I, the e¥uents were equally dis-
tributed and made to £ow through three di¡erent
plastic tanks (1000 L) with black clams (C. £uctifraga)
distributed on the bottom (35 org m  2). In Phase II,
the water treated in each tank with clams £owed into
subsequent tanks (1000 L) containing the benthic
microalgae Navicula sp. at an initial concentration of
50 000 cells mL  1 (Fig. 1). The microalgae were at-
tached to the walls and to arti¢cial substrates (plastic
nets) introduced into the tanks, with a surface area of
1.65 m2.
The untreated e¥uents £owed through the System
B, which had a structure identical to the bioremedia-
tion system (System A), but with no clams or micro-
algae in the 1000 L tanks (Fig. 1).
A third system (System C) was constructed to de-
termine the quality of the inlet estuarine water and
was used as a control of the shrimp rearing in the ef-
£uents. The structure of the System C was identical to
that of Systems A and B, but shrimp were not reared
in the rectangular tanks that received the estuarine
water; also, no clams or microalgae were introduced
into the 1000 L tanks.
Every system was constructed with three repli-
cates with a water £ow of 0.56 L min  1 for every ex-

ESTUARY

Semi-
intensive
culture of
white
shrimp
Effluents

Phase I:
Black clams

Phase II:
Microalgae

Reservoir
SYSTEM A SYSTEM B

Treatment 1 Treatment 2

Figure 1 Scheme of the system used for the bioremediation of shrimp e¥uents (System A) with bivalves (Phase I) and
microalga (Phase II). System B only tested the e¡ect of the pools on the decrease in the water quality parameters.

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Bioremediation and reuse of e¥uents L R Mart|¤ nez-Co¤rdova et al.
perimental unit. The £ow rates of the experimental
systems were controlled by water valves. The hydrau-
lic retention time (HRT) (yh) was 29.7 h for each circu-
lar tank containing clams or microalgae and 184.5 h
for each complete system.
Reuse of treated e¥uents to cultivate white
shrimp
After being treated in Systems A^C, the water was
then reused for rearing shrimp (L. vannamei) in plas-
tic tanks (6 m2) identical to those mentioned above.
Nine pools were used to farm the shrimp in the trea-
ted water. The treated water from System A was dis-
tributed into three pools with a shrimp stocking
density of 25 shrimp m  2 (Fig.1); the water from Sys-
tem B also £owed into the other three culture pools,
and the same was done for System C. Particularly for
System C, the water used was pumped directly from
the estuary, but previously £owed through all the
pools without animals or microalgae. The culture
conditions were the same as mentioned above for
the shrimp culture system.
Analysis of water quality and production
variables
The environmental and water quality variables were
measured in the estuarine inlet water and e¥uents
(before and after bioremediation). The estuarine inlet
water was sampled in the rectangular tanks of Sys-
tem C, which did not have shrimp and received water
directly from the estuary. The raw e¥uents were
measured at the exit of each shrimp culture tank be-
fore the water entered into System A or B. The e¥u-
ents from each system were then analysed as they
exited the tanks of Phase II.
The variables were monitored periodically for tem-
perature, salinity, DO, pH and chlorophyll a twice a
day, using a multisensor YSI 6600 series (Yellow
Springs, OH, USA). Total nitrogen (TN), N-NO2, N-
NO3, TAN and P-PO4, total suspended solids (TSS)
and organic suspended solids (OSS) were all deter-
mined once a week.
The TN was measured using the micro-Kjeldahl
method. The concentration of TAN was evaluated
using the ammonia-salicylate method (Bower &
Holm-Hansen 1980). The nitrite concentration was
determined using the colorimetric method described
by Strickland and Parsons (1972, NitriVer Method). Ni-
trate was determined using the cadmium^copper re-
1418
Aquaculture Research, 2011, 42, 1415–1423
duction method (Wood, Armstrong & Richards 1967),
and the orthophosphates were measured using the
PhosVer 3 method (HACH, Loveland, CO, USA).
To measure the TSS and OSS, 1L of the sampled
water was ¢ltered through GFC 47 mm Whatman ¢l-
ters, which were then washed and dried at 90 1C for
4 h. The di¡erence in weight between the dried ¢lters
before and after ¢ltration was estimated as the TSS.
The OSS were determined by incineration of the dried
¢lters in a mu¥e furnace at 450 1C for 8 h, and then
cooled and weighed. The same process was carried
out with clean seawater (previously ¢ltered and ster-
ilized) from the estuary, which was a basepoint sub-
tracted for the results of TSS and OSS.
The concentration of microalgae cells (Navicula sp.)
was determined by treating samples of water with a
Branson 2210 ultrasonic bath and the concentration
of cells per millilitre was determined in a hematocyt-
ometer.
The shrimp production variables, growth, survi-
val, ¢nal biomass and FCR were evaluated in each of
the pools. The FCR was estimated as the weight of
feed administered/weight gain of the shrimps.
Statistical analysis
A one-way analysis of variance was performed to eval-
uate the production variables of the shrimp cultivated
in the e¥uents from Systems A, B and C, and a post hoc
Tukey test was used to detect signi¢cant di¡erences. A
con¢dence level of 95% was established. The results
are presented as means (standard deviation). To evalu-
ate the water quality variables, a repeated-measures
analysis of variance was performed.
Results
Bioremediation
Signi¢cant di¡erences in some of the water quality
parameters were observed among the raw e¥uents,
the di¡erent Systems of e¥uent processing (A, B or
C) and the inlet estuarine water (Figs 2 and 3).
Most of the parameters monitored for water qual-
ity were signi¢cantly higher in the raw e¥uents and
the e¥uents from System B that were not bioreme-
diated, while lower concentrations of those para-
meters were observed in System A (bioremediated
e¥uents), System C (control) and the Estuary.
The TN increased with time in all the systems,
but signi¢cantly higher values were found in the
raw e¥uents and System B, while no di¡erences were
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Aquaculture Research, 2011, 42, 1415^1423 Bioremediation and reuse of e¥uents L R Mart|¤ nez-Co¤rdova et al.

Total Nitrogen TAN NO2

2.4 0.04 1.2

1.8 0.03 0.9

1.2 0.02 0.6

Concentration (mg·L–1)
0.6 0.01 0.3

0 0 0
1 2 3 4 5 6 7 1 2 3 4 5 6 7 1 2 3 4 5 6 7

NO3 PO4
0.5 0.05
0.4 0.04
0.3 0.03
0.2 0.02
0.1 0.01
0 0
1 2 3 4 5 6 7 1 2 3 4 5 6 7
Time (Weeks)

Figure 2 Concentrations of water quality parameters found throughout time in the raw e¥uents, the bioremediated
e¥uents with bivalve and microalga (System A), the nonbioremediated e¥uents (System B), the estuarine that £owed into
a similar system of tanks but without animals (System C) and the estuarine water (Estuary). Di¡erent letters on the left of
each marker in each graphic indicate signi¢cant di¡erences.

observed among the rest of the Systems (A and C) and Total Suspended Solids
300
the estuarine water (Fig. 2).
250
The concentration of TAN recorded a similar
dynamic in all the treatments through the 200
c Raw Effluents
experimental period; however, the raw e¥uents 150 a System A
and the e¥uents from System B had higher b System B
100 a
concentrations of TAN that those found in System C System C
50 a Estuary
Concentration (mg·L–1)

and the Estuary. In the case of the bioremediated ef-

£uents (System A), the concentrations of TAN were 0
1 2 3 4 5 6 7
signi¢cantly lower than the raw e¥uents but no dif-
ferences were found with regard to the rest of the Organic Suspended Solids
treatments. 50
For nitrite levels, the raw e¥uents and System B 40
showed the highest concentrations during the ex-
periment. Although System A showed results similar 30 c Raw Effluents
to those of System B (P 5 0.069), they were also simi- ab System A
20 b System B
lar to those observed in System C and Estuary a System C
(P40.7). A similar tendency was observed for ni- 10 a Estuary
trates as that for TAN and nitrite concentrations
(Fig. 2). 0
1 2 3 4 5 6 7
The phosphates were also higher in the nonbiore- Time (Weeks)
mediated e¥uents (raw e¥uents and System B) as
Figure 3 Concentrations of suspended solids found through-
compared with the rest of the treatments. System A
out time in the raw e¥uents, the bioremediated e¥uents with
had lower values than the nonbioremediated e¥u-
bivalve and microalga (System A), the nonbioremediated e¥u-
ents, but higher than those of System C and the Estu- ents (System B), the estuarine that £owed into a similar system
ary (Fig. 2). of tanks but without animals (System C) and the estuarine
The TSS were the highest in the raw e¥uents, fol- water (Estuary). Di¡erent letters on the left of each marker in
lowed by those found in System B. Both nonbioreme- each graphic indicate signi¢cant di¡erences.
diated e¥uents had higher values of TSS than
Systems A and C and the estuarine water. Systems A the raw e¥uents and those from System B. No di¡er-
and C and the estuarine water showed statistically si- ences were found among the OSS levels of System A
milar values (Fig. 3). The OSS were also the highest in and the rest of the treatments (Fig. 3).

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Bioremediation and reuse of e¥uents L R Mart|¤ nez-Co¤rdova et al. Aquaculture Research, 2011, 42, 1415–1423

Navicula sp. Table 1 Environmental variables in the microcosm culture

100000
of shrimp in the bioremediated e¥uents (System A), the non-
80000 bioremediated e¥uents (System B) and the estuarine water
Cells·mL–1

(System C)
60000

40000 System A System B System C

20000 Temperature ( 1C) 26.1  1.1a 26.2  1.2a 25.9  0.9a

Salinity (%) 41.2  2.1a 41.5  2.0a 41.0  1.9a
0
1 2 3 4 5 6 7 DO (mg L  1) 5.6  0.5a 5.7  0.6a 5.9  0.6a
Time (Weeks) pH 8.2  0.3a 8.2  0.2a 8.3  0.2a
NO3 (mg L  1) 0.30  0.06a 0.41  0.05b 0.28  0.05a
Chione fluctifraga NO2 (mg L  1) 1.04  0.55a 1.51  0.76b 1.01  0.62a
100 TAN (mg L  1) 0.68  0.35ab 1.05  0.6b 0.60  0.32a
PO4 (mg L  1) 0.16  .04a 0.21  .09a 0.15  0.06a
80
Survival (%)

Di¡erent letters in the same row indicate signi¢cant di¡erences

60 (Po0.05).
DO, distilled water; TAN, total ammonia nitrogen.
40

20 Table 2 Production parameters of white shrimp farmed at

0
microcosms in the bioremediated e¥uents (System A), the
1 2 3 4 5 6 7 nonbioremediated e¥uents (System B) and the estuarine
Time (Weeks) water (System C)

Figure 4 Concentration of the benthic microalga (Navi-

System A System B System C
cula sp.) and survival of clams (Chione £uctifraga) during
the experiment. Final weight (g) 14.2  1.3a 13.8  1.2a 14.0  1.6a
Growth rate 1.15  0.04a 1.10  0.05a 1.12  0.06a
(g week  1)
In terms of e⁄ciency, System A with clams and mi-
Survival (%) 89.2  2.1a 75.1  3.6c 82.1  3.8b
croalgae removed 17.3% of TN, 24.5% of TAN, 19.2% of
Final biomass 3166  206a 2610  280b 2874  205ab
nitrites, 13.5% of nitrates, 21.6% of phosphates, 22.3% (kg ha  1)
of TSS and 23.2% of OSS from the raw e¥uents. System FCR 1.47  0.06a 1.73  0.05b 1.51  0.05a
B showed a removal e⁄ciency of o4% of TN, nitrites,
Di¡erent letters in the same row indicate signi¢cant di¡erences
nitrates and phosphates, while the removal of TAN,TSS (Po0.05).
and OSS was 10.2%,10.1% and 8.6% respectively. FCR, feed conversion ratio.
The concentration of the benthonic microalgae re-
mained at levels above the 50 000 cells mL  1, during mental variables: temperature, salinity, DO, pH and
the ¢rst 5 weeks of the experiment, but declined dur- PO4 (Table 1). However, higher values of TAN, NO2
ing the last week (Fig.4). By the last week, the presence and NO3 were observed in the tanks where shrimp
of Navicula sp. was replaced in part by an unidenti¢ed from System B were reared.
diatom. The chlorophyll levels were signi¢cantly Regarding the production parameters of the
higher in Phase II of System A (23.5  3.8 mg m  3), shrimps farmed in di¡erent types of e¥uents, the ¢-
which had the benthonic microalgae, followed by nal weight and the growth rate were statistically si-
Phase II from Systems B (7.0  2.6 mg m  3) and C milar among all the treatments (Systems A, B and C).
(4.0  2.7 mg m  3) respectively. The survival of the However, the highest survival was found in shrimps
clams was constant during the entire experiment, reared in the bioremediated e¥uents (System A), fol-
with an average survival above 90% in every week lowed by those reared in estuarine water (System C),
(Fig. 4); as adult clams were used for the experiment, while the lowest survival was observed in the
the growth during the experimental period was not shrimps cultivated in nonbioremediated e¥uents
signi¢cant (0.5^1.0 mm). (System B) (Table 2).
The highest biomass was also achieved in System
A and the lowest in System B, while no di¡erences
Shrimp rearing in e¥uents
were observed among System C and the rest of the
No signi¢cant di¡erences were detected among the treatments. The biomass of shrimp from System A
treatments with respect to the following environ- was 21% higher than that of System B (Table 2).

1420 r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 1415^1423

Aquaculture Research, 2011, 42, 1415^1423
Finally, FCRs were lower in Systems A and C com-
pared with those obtained for the shrimps reared in
System B (Table 2).
Discussion
Shrimp rearing at a semiintensive scale is activity
practice capable of exerting a signi¢cant impact on
the environment. Some of the parameters of water
quality increased by 100% or more from the estuar-
ine water to the raw e¥uents. Although this was de-
monstrated at an experimental scale, similar results
have been observed in commercial farms (Jackson,
Preston & Thompson 2004). Jackson et al. (2004) stu-
died the discharge nutrient loads at di¡erent shrimp
farms, ¢nding concentrations of TN and TSS as high
as 3 and 200 mg L  1, respectively; similar results
were observed in this experiment.
The bioremediation system appeared to have a
moderate e⁄ciency, as nitrogenous compounds and
the phosphates were lower after the e¥uent £owed
through the pools with clams and the benthonic mi-
croalgae. Some authors have documented that the
presence of bivalves and microalgae can decrease
the concentration of di¡erent compounds that con-
taminate receiving ecosystems. For instance, Her-
nandez, Bashan and Bashan (2006) found a P
removal e⁄ciency of around 25% for Chlorella spp.
alone, and up to 72% for Chlorella spp. co-immobi-
lized with Azospirillum braziliense. In addition, Jones,
Dennison and Preston (2001) evaluated a multi-
phase system (sedimentation, ¢lter feeders and mi-
croalgae) to treat shrimp e¥uents and achieved an
overall improvement in water quality as follows: TSS

(12%), TN (28%), P (14%), NH1 4 (76%), NO3 (30%),

PO4 (35%), bacteria (30%) and chlorophyll a (0.7%);
however, the HRT of this system was lower than that
observed in our experiment, which suggests that the
e⁄ciency of our system was lower that of Jones et al.
(2001). Also, the density of bivalves they used was
much higher, although the retention time used in
our system corresponded to a daily water exchange
similar to that used in commercial farms (10^20%).
The bioremediation e⁄ciency in the above-men-
tioned experiments as well as in our experiment was
estimated by measuring the number of nutrients
removed from the raw e¥uents. The results suggest
that the quality of water was signi¢cantly improved
after being treated by the bioremediation system (A);
however, System B, which did not have clams or mi-
croalgae, also showed some e⁄ciency in removing
TAN,TSS and OSS. These results may be attributed to
r 2010 Blackwell Publishing Ltd, Aquaculture Research, 42, 1415^1423
Bioremediation and reuse of e¥uents L R Mart|¤ nez-Co¤rdova et al.
the sedimentation in the pools. Hence, it can be hy-
pothesized that the e⁄ciency of System A in remov-
ing nitrogenous metabolites and phosphates can be
attributed to the presence of Navicula sp., while the
removal of suspended solids may be caused in part
by the presence of clams and by sedimentation in
the tanks (sediments were observed in the treatment
pools of Systems A and B, although they were not
measured). Vymazaj (1988) found that microalgae
species such as Navicula sp. were capable of removing
nutrients from polluted streams with a maximum ef-
¢ciency of 80% and 70% for ammonium and ortho-
phosphates respectively. Navicula sp. has been used as
part of bio¢lms to improve the water quality, due to its
ability to remove nitrogenous compounds and phos-
phates (Thompson, Abreu & Wasielesky 2002). Re-
garding suspended solids, the presence of bivalves
and the use of tanks as sedimentation units have
been shown to decrease the concentration of sus-
pended solids and TN from aquaculture e¥uents
(Jones et al. 2001; Bernal-Jaspeado 2006; Li, Veilleux
& Wikfors 2009).
The greater e⁄ciency of System A in removing OSS
than TSS could be explained by the ¢ltration activity
of the clams; in this regard, it has been demonstrated
that bivalves preferably ingest organic and reject in-
organic materials (Newell & Jordan 1983).
Although the bioremediation system with C. £ucti-
fraga and Navicula sp. had acceptable e⁄ciency, the
concentrations of the microalgae decreased during
the last week. This decrease may be attributed to the
high turbidity observed in System A during the last 2
weeks, caused by the increase in TSS and OSS. It has
been reported that the abundance of some microal-
gae species such as Navicula sp. depends on the tur-
bidity and the concentration of suspended solids
(Unrein & Vincour 1999). Some alternatives to solve
this problem might be to increase bivalves in Phase I
or include a sedimentation tank, as suggested by
Jones et al. (2001) to decrease the concentration of
suspended solids. Although Navicula sp. levels de-
creased by the last week, the nitrogenous metabolites
continued decreasing in System A, which may be at-
tributed to the remaining concentration of Navicula
sp. and the unidenti¢ed diatom that was predomi-
nant during the last few days.
As a ¢rst approach, it was observed that the
bioremediation system had lower e⁄ciency than
those using macroalgae, in terms of nutrient removal
and biomass production (Xu, Fang & Wei 2008;
Marinho-Soriano, Nunes, Carneiro & Pereira 2009;
Mart|¤ nez-Porchas et al. 2010). However, it is important
1421
Bioremediation and reuse of e¥uents L R Mart|¤ nez-Co¤rdova et al.
to continue studying the ability of C. £uctifraga and
Navicula sp. as potential bioremediators of shrimp
(or ¢sh) e¥uents, at di¡erent densities and using dif-
ferent system designs, to achieve a better e⁄ciency.
Regarding the production parameters of the
shrimp reared on the three systems, it was observed
that the bioremediated e¥uents (System A) were
very suitable for the culture of white shrimp.
The growth rates were similar to or higher than the
0.9^1.0 g week  1, reported as commercially feasible
(Mart|¤ nez-Co¤rdova 1999). The productive response of
the shrimp reared in the bioremediated e¥uents was
better than that observed in those cultivated in e¥u-
ents from Systems B and C. The di¡erence in survival
and biomass among the shrimps reared in bioreme-
diated e¥uents (from System A) and those reared
in the nonbiormediated e¥uents (System B) may be
attributed to the improvement in the water quality.
Although none of the water quality parameters
reached the lethal concentration (LC50) in the non-
bioremediated e¥uents, they were almost twofold
higher than those from the bioremediated e¥uents.
Di¡erent authors have documented that the chronic
exposure of penaeid shrimps to high concentrations
of nitrogenous compounds and suspended solids can
diminish their growth and food intake (Frias-Esperi-
cueta, Harfush-Melendez & Paez-Osuna 2000; Ray,
Lewis, Browdy & Le¥er 2009). Moreover, the slight
increase in the productive response of shrimps from
System A as compared with those reared in System C
could be attributed to the higher survival and to the
presence of OM in System A, such as bio£ocs and mi-
croalgae (Navicula sp. and other diatoms), which
could be an alternative source of food.
The biomass obtained in Systems A and C was
higher than the mean reported in most semiintensive
farms of the region. The FCRs in the same treatments
are considered to a pro¢table value for commercial
purposes (Juarez 2008). The results suggest that the
shrimp can thrive in bioremediated e¥uents, which
indicates that the treated water (by clams and micro-
algae) can be reused by recirculation. This practice
would reduce the environmental impact caused by
the massive discharges of shrimp aquaculture.
Furthermore, the black clam have commercial value
and may represent and extra economical income for
farmers (Mart|¤ nez-Porchas et al. 2010).
It can be concluded that the bioremediation system
was moderately e⁄cient in removing nutrients and
solids (TSS and OSS) from shrimp aquaculture e¥u-
ents. Also, the tanks themselves are useful due to
the sedimentation activity.
1422
Aquaculture Research, 2011, 42, 1415–1423
It is necessary, however, to improve System A with
some modi¢cations in the design. The production
parameters obtained in the present study strongly
suggest that the bioremediated e¥uents can be used
for farming white shrimp without a negative e¡ect on
its survival and growth.
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