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Aquaculture 321 (2011) 130–135

Contents lists available at SciVerse ScienceDirect

Aquaculture
journal homepage: www.elsevier.com/locate/aqua-online

Effect of calcium hydroxide, carbonate and sodium bicarbonate on water quality and
zootechnical performance of shrimp Litopenaeus vannamei reared in bio-flocs
technology (BFT) systems
Plínio S. Furtado, Luís H. Poersch, Wilson Wasielesky Jr. ⁎
Marine Station of Aquaculture, Institute of Oceanography, Federal University of Rio Grande, C. P. 474, Rio Grande (RS), CEP 96201-900, Brazil

a r t i c l e i n f o a b s t r a c t

Article history: Litopenaeus vannamei (the Pacific white shrimp) is the most commonly reared species in super-intensive bio-
Received 28 March 2011 floc technology (BFT) without water renewal. In BFT, the pH may decrease due to the reduction of alkalinity
Received in revised form 23 August 2011 and the increase of dissolved carbon dioxide. This study evaluated the effects of calcium hydroxide, sodium
Accepted 25 August 2011
carbonate and bicarbonate in maintaining water quality during the cultivation of L. vannamei in BFT. The ex-
Available online 8 September 2011
periment was conducted using juveniles stocked in 150-L 12 tanks at a density of 333 shrimp/m 3. There were
Keywords:
four treatments with three replicates each: T1 — Na2CO3, T2 — Ca(OH)2, T3 — NaHCO3 and T4 — control. For
Litopenaeus vannamei pH correction, alkalinity and both pH and alkalinity corrections, we used sodium carbonate (0.06 g/L), sodi-
pH um bicarbonate (0.20 g/L) and hydrated lime (0.15 g/L), respectively. Significant physical, chemical and bio-
Alkalinity logical differences (P b 0.05) were detected among treatments. Control shrimps showed lower growth
Hydrated lime performance (P b 0.05) than shrimps in other treatments. Hydrated lime and sodium bicarbonate appeared
Bioflocs effective in supplementing alkalinity, whereas the soda ash did not. We verified the effectiveness of sodium
carbonate in raising pH levels and assisting in supplementing alkalinity. This study demonstrates that the re-
sults obtained in the control treatment levels of alkalinity and pH decrease during rearing, and that CO2 levels
would be increased in super-intensive systems with bioflocs without water renewal.
© 2011 Elsevier B.V. All rights reserved.

1. Introduction According to Samocha et al. (2007), the addition of organic carbon

The aquaculture without water exchange, under high stocking
density, with strong aeration and biota (predominantly, aerobic
and heterotrophic), formed by microbial flocs or bio-flocs is known
as ZEAH (Zero Exchange Aerobic Heterotrophic) or, more recently,
as bio-flocs technology (BFT) (Avnimelech, 2007; De Schryver et
al., 2008). Relative to conventional culture systems, these systems
are considered environmentally friendly due to decreased water
use, reduced risk of introduction and spread of pathogens, optimiza-
tion of cultivable area and the opportunity to use areas away from
environmentally protected coastal zones (McIntosh et al., 2000,
Moss et al. 2001; Samocha et al., 2001, Burford et al., 2003). Addi-
tionally, it is possible to increase the productivity in these systems
because the diet is improved through the natural productivity (bio-
flocs), making diets lower in protein possible and, consequently,
reducing the quantity of fishmeal (commonly used in formulation
of diets for fish and shrimp) required (Ballester et al., 2010; Samocha
et al., 2004).
⁎ Corresponding author. Tel.: + 55 53 3236 8131; fax: + 55 53 3236 8042.
E-mail address: manow@mikrus.com.br (W. Wasielesky).
sources, such as sugar cane molasses, can be used to prevent increases
in total ammonium nitrogen and nitrite concentrations in BFT systems.
Additionally, Wasielesky et al. (2006a) concluded that the suspended
particulate matter in Litopenaeus vannamei BFT culture systems can sig-
nificantly improve feed conversion rate reducing production cost. Be-
cause there is no water exchange in these systems, alkalinity tends to
decrease during culture. These systems present photoautotrophic, che-
moautotrophic and predominantly heterotrophic bacteria. According to
Ebeling et al. (2006), for each gram of ammonium nitrogen converted
into heterotrophic microbial biomass, 4.71 g of dissolved oxygen,
3.57 g of alkalinity and 15.17 g of carbohydrate are consumed and
8.07 g of microbial biomass and 9.65 g of carbon dioxide are produced.
The alkalinity of a liquid refers to the total concentration of titratable
bases able to neutralize hydrogen cations. Bicarbonate (HCO3−) and car-
bonate (CO3−2) are the main bases responsible for the total alkalinity,
which is expressed in equivalents of calcium carbonate (mg CaCO3/L).
The total alkalinity is the buffering capacity i.e., the capacity to maintain
the acid–base equilibrium: CO2+H2O ↔ H2CO3 ↔ H+ + HCO3−↔2H++
CO3−2 (Barbieri and Ostrensky, 2002; Vinatea, 1997). According to Van
Wyk and Scarpa (1999), L. vannamei needs an alkalinity higher than
100 mg CaCO3/L for ideal development. The same authors affirm that
high alkalinity reduces the daily fluctuation in pH. Vinatea et al.
(2010), working with L. vannamei in BFT system at densities below

0044-8486/$ – see front matter © 2011 Elsevier B.V. All rights reserved.
doi:10.1016/j.aquaculture.2011.08.034
 Author's personal copy
P.S. Furtado et al. / Aquaculture 321 (2011) 130–135
610 shrimp/m2, used pure sodium bicarbonate (0.12 g/L) to increase
the alkalinity when it dropped below 60 mg CaCO3/L. Several other
studies reported the reduction of alkalinity with the BFT culture system
(McIntosh, 2001; Ray et al., 2010).
In culture systems, the marine shrimp develop best at a pH between
7.0 and 9.0 (Van Wyk and Scarpa, 1999). The pH influences almost all
chemical reactions and phenomena that occur in the water, and it also
influences the physiological conditions of the shrimp (Lemonnier et
al., 2004). Environments with a pH below 6.5 or above 9.5 can reduce
the growth of Penaeidae. Additionally, a pH between 7.0 and 9.0 favors
the development of heterotrophic and nitrifying bacteria (Chen et al.,
2006; Van Wyk and Scarpa, 1999).
According to McIntosh (2001), a high stocking density causes an
increase in the respiratory rate and increased levels of CO2, and
thus, a reduction in the pH. L. vannamei reared at Belize Aquaculture
Ltda. (BAL) did not negatively react to pH of 6.8 to 7.3. On the other
hand, Wasielesky et al. (2006b), showed that the reduction in the
pH to below of 7 in a BFT system does affect negatively the growth
of L. vannamei. Thus, in order to improve the techniques of shrimp
culture in BFT systems, it is important to carry out experiments that
evaluate the effects of alkalinizing compounds on the water quality
and zootechnical performance of L. vannamei in systems with BFT.
2. Material and methods
2.1. Biological material and experimental design
This study was conducted between February and May 2009 at
the Marine Station of Aquaculture (EMA), Institute of Oceanogra-
phy, Federal University of Rio Grande, Rio Grande do Sul — Brazil.
L. vannamei juveniles were reared in a BFT system with densities
below 300 shrimp/m 2 until the animals reached a mean weight of
6 g. Thereafter, shrimp were transferred to the experimental units
(150 L), at densities of 50 shrimp/tank (equivalent to 1 shrimp/3 L
or 333 shrimp/m 3). The experimental design consisted of four treat-
ments with three replicates each: T1 – Na2CO3 – in which the pH
was maintained above 7.5 through the use of sodium carbonate;
T2 – Ca(OH)2 – in which the pH was maintained above 7.5 and
the alkalinity above 100 mg CaCO3/L through the use of calcium hy-
droxide; T3 – NaHCO3 – in which the alkalinity was maintained
above 100 mg CaCO3/L through the use of sodium bicarbonate;
and T4 – control – in which neither the pH nor the alkalinity was
corrected during the 60 day experiment. The shrimp were fed a
commercial diet (Guabi/38 Active — 38% crude protein, 8% lipid,
2 mm pellet size) through the use of feeding trays (at 07:00 and
17:00). The initial feeding rate was based on Jory et al. (2001),
with adjustments according to the consumption observed in a peri-
od of 24 h and results of mean weight of shrimp caught weekly
through biometrics. The percentage of leaching was verified for
the subsequent quantification of feed consumption.
2.2. Correction of alkalinity and pH
The correction of pH levels and alkalinity during the study oc-
curred when pH (≤7.5) and alkalinity (≤100 mg CaCO3/L). The effi-
ciency of the chemical compounds used to correct the pH and
alkalinity was analyzed in seawater with microbial flocs and without
shrimp. The tests were performed, with two replicates, in plastic
tanks (2 L of useful volume) with one airstone each to provide aera-
tion. The pH and alkalinity were measured before use (pH initial 7.0
and alkalinity initial 100 mg CaCO3/L), 1 h after use and 24 h after
the use of the chemical compounds. The tested chemical compounds,
as well as their dosage, purpose and pH and alkalinity increase levels,
are shown in Table 1. Sodium carbonate (Na2CO3) is water-soluble,
has a molecular weight of 106 g, 94% neutralization power (PN),
96% reactivity rate (ER) and 95% relative power of total neutralization
131
Table 1
Chemical compounds used for the correction of pH and alkalinity in the culture of
L. vannamei in treatments with Na2CO3 for pH, NaHCO3 for alkalinity and Ca(OH)2 for
both corrections.
Correction Chemical Concentration Increase level
compound (g/L)
pH Alkalinity
(mg CaCO3/L)
pH Na2CO3 0.06 0.7 ± 0.1 20 ± 10
pH and Ca(OH)2 0.15 0.8 ± 0.15 110 ± 10
alkalinity
Alkalinity NaHCO3 0.20 0.25 ± 0.05 100 ± 10
The data correspond to the mean of 2 replicates ± standard deviation.
(PRNT). Sodium bicarbonate (NaHCO3) is water-soluble, has a molec-
ular weight of 84 g, 56% PN, 97% ER and 55% PRNT. Calcium hydroxide
(Ca(OH)2) is water-soluble, has molecular weight of 74 g, 132% PN,
62% ER and 81% PRNT. The quantities of the chemical compounds re-
quired to correct pH and alkalinity were calculated according to the
equations below:
K ¼ Nf −Ni ð1Þ
where K is the level to be increased (pH or mg CaCO3/L); Nf is desired
level (pH or mg CaCO3/L); and Ni is current level (pH or mg CaCO3/L).
Y ¼ ðW  K Þ=Z ð2Þ
where Y is the chemical compound concentration to be used (g/L); W
is concentration used in the test (g/L); and Z is the capacity of W in
increasing the parameter (pH or alkalinity).
Q ¼ Y V ð3Þ
where Q is the chemical compound quantity to be used (g); Y is con-
centration of the chemical compound (g/L); and V is the useful vol-
ume of the tank (L).
2.3. Organic fertilization
The microbial flocs were inoculated from a culture, which the
shrimp were originated, at a rate of 30% of the experimental units' use-
ful volume and, during the experiment, no water exchange was per-
formed. It was necessary to add organic fertilization to adjust the
carbon:nitrogen (15:1) ratio. The addition of sugar cane molasses
(37.46% C and 0.57% N) into carbon source was based on Ebeling et al.
(2006) and Avnimelech (1999) who determined that 6 g C is needed
to convert 1 g total ammonium nitrogen (TA-N) into bacterial biomass.
The addition of sugar cane molasses was made when the concentrations
of TA-N were equal or greater than 0.8 mg/L.
2.4. Physical and chemical parameters and zootechnical performance
The photoperiod was 12 h of dark followed by 12 h of light (artificial
luminosity of 50 lx). The temperature of the water was maintained at
27 °C through the use of heaters with a thermostat (VisiTherm —
Italy). Aeration was provided through hoses and airstones. The salinity
was maintained at 33‰ by adding, when necessary freshwater.
Temperature, pH and dissolved oxygen were measured twice daily
(morning and afternoon) with a pH-meter (WTW pH 315i, Germany)
and an oximeter (WTW OXI-315i, Germany). The salinity was mea-
sured every 3 days with an optical refractometer (Atago, 103, optical
refractometer ±1 ppt, Japan). The concentrations of TA-N and nitrite
(NO2-N) were measured three times a week, according to UNESCO
(1983) and Bendschneider and Robinson (1952), respectively. The
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132 P.S. Furtado et al. / Aquaculture 321 (2011) 130–135

dissolved carbon dioxide (CO2) was calculated with the software CO2 450
Analysis Salt® (Timmons and Ebeling, 2010). The alkalinity was ana-

Volume Microbial Flocs (mL/L)

400
lyzed three times a week, and the orthophosphate (PO43–P) and nitrate
350
(NO3–N) were measured every 7 days according to APHA (1998) and
Aminot and Chaussepied (1983), respectively. Total suspended solids 300
(TSS) were determined every 7 days with gravimetry by filtering sam-
250
ples of 20 mL in glass-fiber filters (GF 50-A) (Strickland and Parsons,
1972). TSS values were estimated by the difference between final and 200
initial weight of each filter (AOAC, 2000). The volume of microbial
150
flocs (MFV) was obtained through the use of Imhoff cones (Avnimelech,
2007). 100
Initially and every 7 days after, a sample of 10 shrimp from each
50
tank was weighed with a precision scale (± 0.01 g SD-Marte®). At
the 60th experimental day all live shrimp were quantified and 0
1 11 21 31 41 51 61
weighed for the evaluation of growth and survival in each treatment.
Days
The following metrics were determined: survival (S; %) = [(initial
T1 T2 T3 T4
shrimp n o − dead shrimp n o)/initial shrimp n o] × 100; final weight
(FW; g) = Σ final weight of live shrimp/total shrimp n o; weight gain Fig. 1. Mean concentrations of 3 replicates of the volume of microbial flocs in the culture
(WG; g) = final weight − initial weight; weight gain/week (WG/W; of Litopenaeus vannamei juveniles in the different experimental conditions (T1—Na2CO3,
g/week) = weight gain/culture weeks; final biomass (FB; g) = Σ final T2—Ca(OH)2, T3—NaHCO3, T4—control) during 60-day treatments.

weight of live shrimp; feed conversion rate (FCR) = (offered diet

− leaching)/weight gain; and productivity (P; kg/m 3) = (final bio-
mass − initial biomass) × 6.66.
2.5. Statistical analysis
After verifying the normality and homoscedasticity of the data, we
performed a one-way analysis of variance (ANOVA) to verify if there
were significant differences among treatments. When it was detected
(P b 0.05), the Tukey test of means comparison was used. The percent-
age values were transformed (square root arcsine) prior to the analysis
(Zar, 1996). We used the software package STATISTICA 7.0 (StatSoft Inc.
2004, Tulsa, Oklahoma, USA) for these calculations.
with the highest mean recorded in T2, followed by T1, T3 and T4.
The alkalinity showed significant difference among treatments
(P b 0.05), with the highest mean value found in T2, followed by T3,
T1 and T4. The T4 showed the highest dissolved CO2 means
(P b 0.05), followed by T3, T1 and T2. The progression of the alkalinity
and pH mean values are shown in Fig. 2. The zootechnical perfor-
mance parameters are shown in Table 3. The FW and WG means
were significantly higher in T1 (P b 0.05), with T4 having the lowest
A
250
Alkalinity (mg CaCO3/L)

3. Results 200

3.1. Physical and chemical parameters and zootechnical performance 150

The physical and chemical parameters are shown in Table 2. The

100
MFV means were significantly higher (P b 0.05) for T1, T2 and T3.
The progression of the MFV means is shown in Fig. 1. The PO43–P
mean values were significantly higher (P b 0.05) for T4, followed by 50
T1, T3 and T2. The alkalinity, pH and CO2 mean values are shown in
Table 2. The pH differed significantly (P b 0.05) among treatments, 0
1 11 21 31 41 51 61
Days
Table 2 T1 T2 T3 T4
Mean values ± standard deviation of the water quality parameters during culture
(60 days) of L. vannamei in different experimental treatments1.
B
Parameters T1—Na2CO3 T2—Ca(OH)2 T3—NaHCO3 T4—control 9.0

Temperature 27.05 ± 2.3 27.25 ± 2.2 27.12 ± 2.1 27.0 ± 2.0 8.5
(°C)
Salinity (‰) 33.50 ± 1.5 33.50 ± 1.5 33.5 ± 1.0 33.0 ± 1.5
8.0
DO2 (mg/L) 5.0 ± 0.3 4.9 ± 0.4 5.1 ± 0.3 5.1 ± 0.3 7.5
CO2 (mg/L) 1.06 ± 0.4A 1.37 ± 0.3A 5.38 ± 6.6B 12.81 ± 16.1C
pH

pH 7.77 ± 0.5AB 7.90 ± 0.5A 7.5 ± 0.46B 6.86 ± 0.9C 7.0

Alkalinity 100 ± 33.9C 162.2 ± 44.9A 144.5 ± 30.1B 78 ± 47.2D
6.5
(mg CaCO3/L)
MFV (mL/L) 108.47 ± 87.6B 112.22 ± 112.5B 121.72 ± 95.9B 67.37 ± 56.1A 6.0
TSS (mg/L) 850 ± 470 890 ± 628 860 ± 450 870 ± 462
TA-N (mg/L) 0.3 ± 0.4 0.4 ± 0.7 0.4 ± 0.5 0.6 ± 0.7 5.5
NO2–N (mg/L) 3.1 ± 5.5 3.5 ± 5.9 4.0 ± 6.5 4.3 ± 7.0
5.0
NO3–N (mg/L) 57.5 ± 33.9 59.5 ± 33.9 60.53 ± 35.5 54.5 ± 34.5 1 11 21 31 41 51 61
PO43–P (mg/L) 6.68 ± 2.4AB 5.0 ± 1.3A 6.11 ± 2.8AB 8.2 ± 1.3B Days
1
The data correspond to the mean of 3 replicates ± standard deviation. Different T1 T2 T3 T4
superscripts in the same row indicate that the means, significantly, differ (P b 0.05).
Dissolved oxygen (DO2), dissolved carbon dioxide (CO2), volume of microbial flocs Fig. 2. Mean concentrations of the alkalinity (A) and pH (B) in the culture of Litopenaeus
(MFV), total suspended solids (TSS), total ammonium nitrogen (TA-N), nitrite (NO2–N), vannamei juveniles in the different experimental conditions (T1—Na2CO3, T2—Ca(OH)2,
nitrate (NO3–N) and orthophosphate (PO43–P). T3—NaHCO3, T4—control) during 60-day treatments.
 Author's personal copy

P.S. Furtado et al. / Aquaculture 321 (2011) 130–135 133

Table 3 initially bactericidal due to the stress caused by the pH increase and
Zootechnical parameters of L. vannamei juveniles reared during 60-day experimental the modification in the CO2–HCO3−–CO3−2 equilibrium.
treatments with correction of pH (T1—Na2CO3), pH and alkalinity (T2—Ca(OH)2),
alkalinity (T3—NaHCO3) and without correction of pH and alkalinity (T4—control).
In T2, we used dosages of up to 0.16 g/L of calcium hydroxide;
however there was no inhibition or decontrol of the nitrifying bacte-
Parameters T1—Na2CO3 T2—Ca(OH)2 T3—NaHCO3 T4—control ria groups because the nitrification was similar to that of T1 and T3.
Survival (%) 83.3 ± 3.0 85.0 ± 7.0 80.0 ± 2.7 80.0 ± 2.8 NO3–N accumulated during the experiment in all treatments, and
Initial weight (g) 5.6 ± 0.9 5.8 ± 0.9 5.6 ± 0.7 5.8 ± 1.1 the mean concentrations remained below 60 mg/L; however, all
Final weight (g) 15.0 ± 1.3A 14.3 ± 1.4B 14.2 ± 1.6B 12.0 ± 1.5C
treatments showed maximum values of above 60 mg/L. Kuhn et al.
Weight gain (g) 9.3 ± 1.2A 8.4 ± 1.5AB 8.5 ± 0.1B 6.2 ± 0.2C
Final biomass (g) 630.0 ± 61.1A 615.7 ± 9.3A 570.8 ± 7.2B 480.0 ± 17.8C (2010) reported the negative effect of NO3–N on the L. vannamei
Weight gain/week 1.1 ± 0.2A 1.0 ± 0.1A 1.0 ± 0.1A 0.7 ± 0.1B growth and survival when in concentrations N220 mg/L and at low
(g/week) salinities (≈ 11‰). In T4 there was a reduction in the NO3–N values
Feed conversion rate 1.4 ± 0.2A 1.4 ± 0.1A 1.5 ± 0.1A 3.0 ± 0.3B during the last 20 days. This is likely a result of denitrification, in
Productivity (kg/m3) 2.3 ± 0.1A 2.2 ± 0.1A 1.8 ± 0.1B 1.3 ± 0.5C
which anaerobic bacteria convert nitrate into nitrogen gas (N2; Van
The data correspond to the mean of 3 replicates ± standard deviation. Different Wyk and Scarpa, 1999).
superscripts in the same row indicate that the means, significantly, differ (P b 0.05).
The PO43–P concentrations remained below the 40 mg/L obtained by
Ray et al. (2010), but were higher than the values in other studies in BFT
mean values. The FB means were significantly higher in T1 and T2 systems (Krummenauer, 2008; McIntosh et al., 2000). The accumula-
(P b 0.05). The WG/W and FCR mean values were significantly best tion of phosphorus is due to the non-ingested feed and excreta de-
in T1, T2 and T3. The P means were significantly higher (P b 0.05) in composition, which favors eutrophication (Peñaflorida, 1999). This
T1 and T2, followed by T3 and T4. accumulation does not directly affect the development of shrimp,
but it may cause favorable conditions for the proliferation of filamen-
tous cyanobacteria, which can obstruct the shrimp gills and produce
3.2. Correction of alkalinity and pH
harmful toxins (Wasielesky et al., 2006b). Silva (2009), evaluating the
nitrogen and phosphorus dynamics in L. vannamei and Farfantepenaeus
The correction of alkalinity and pH was performed at values of
paulensis culture in BFT system, affirms that the accumulation of phos-
100 mg CaCO3/L and ≤7.5, respectively. The T2 had the pH and alka-
phorus, and the microbial floc inability to retain great quantities of it,
linity corrected for three times: on the 20th, 39th and 56th experi-
make excess phosphorus removal necessary.
mental days (Fig. 2A). The T3 had the alkalinity corrected three
The TSS values increased during the experimental period in all
times: on the 20th, 37th and 50th experimental days (Fig. 2A). The
treatments, and the means were higher than the 643 mg/L reported
T1 had the pH corrected five times: after the 27th experimental day
by Avnimelech (2007). The same pattern was verified for the MFV,
and, thereafter, every 8 days (Fig. 2B). The T4 had no correction of
with concentrations higher than 100 mL/L, except in T4, which showed
pH and alkalinity resulting in the reduction of both (Fig. 2A and B).
a mean of 67 mL/L. Conditions of pH less than 7 and alkalinity below
100 mg CaCO3/L probably contributed to the reduced development of
4. Discussion the microbial flocs observed in T4. These values are high, as Samocha
et al. (2007) recommend concentrations of TSS and MFV up to
In this study the lowest temperature was above 23.6 °C, and the 500 mg/L and 10 mL/L, respectively.
mean was 27 °C. Temperature remained within the recommended According to Vinatea et al. (2010), there is an inverse relationship
range for the best growth and survival of these species juveniles between suspended particulate matter and photosynthesis, where
(Ponce-Palafox et al., 1997). Ponce-Palafox et al. (1997) obtained high turbidity reduces the light penetration and, thus, photosynthe-
the best survival and growth of L. vannamei juveniles at salinities be- sis. The MFV may have been influenced by the low luminosity
tween 33 and 40‰. The mean salinity in this experiment was of (50 lx) of the experimental room because microbial populations are
33.5‰, which is within the ideal range. According to Van Wyk and more stable and less dependent on luminosity (Avnimelech, 2006).
Scarpa (1999), dissolved oxygen concentration should be higher In this study, it is possible that the high values of TSS and MFV nega-
than 5.0 mg/L, in this study, although the values were slightly below tively affected the zootechnical performance through the obstruction
the recommended range. of the shrimp gills. When the TSS and MFV exceed the levels recom-
The TA-N from the excretion and decomposition of organic matter mended by Samocha et al. (2007), as they did here, measures must
may affect the performance at concentrations above 3.95 mg/L and at be taken to reduce the MFV. Ray et al. (2010) verified that in tanks
salinity of 35‰, causing mortality (Lin and Chen, 2001). The NO2–N is where the TSS were removed, there was a reduction by 59% in TSS,
a compound that accumulates in the culture. At salinity of 35‰, values 60% in NO3–N, 61% in PO43–P and an increase of 33% in alkalinity. To
of 25.7 mg/L and above can reduce L. vannamei growth (Lin and Chen, reduce the TSS, NO3–N and PO43–P hydro-cyclones and clarifiers can
2003). NO3–N is the final product of ammonia nitrification, and it is be used.
the nitrogen compound that is the least harmful to Penaeidae, being The dissolved CO2 in BFT systems has not been well studied. Howev-
safe above 60 mg/L (Van Wyk and Scarpa, 1999). In this study, the TA- er, in recirculating systems for fish culture it has already been investi-
N concentrations were within the ideal range for this juvenile culture gated by several researchers (Moran, 2010; Summerfelt et al., 2000).
(Lin and Chen, 2001). During the experiment, there was a great fluctu- The CO2 is toxic to aquatic organisms because it reduces the capacity
ation in the TA-N concentration of T4. This suggests the need for more of the hemolymph to transport oxygen, thereby acidifying the hemo-
organic fertilization. lymph and inducing metabolic stress. Van Wyk and Scarpa (1999)
The nitrite values remained within adequate levels (below 25.7 mg/L found that values of CO2 below 5 mg/L are ideal, 20 mg/L are acceptable,
at 35‰) for the L. vannamei juvenile culture (Lin and Chen, 2003), and 20–60 mg/L are not lethal, but cause interferences in the CO2 exchange
there were no significant differences among treatments (P N 0.05). The in the shrimp gills, and N60 mg/L may be fatal for Penaeidae.
initial concentration of 10 mg/L was increased due to the values and The accumulation of CO2 in BFT systems occurs through the respi-
the already structured microbial chain of the bio-floc inoculums. Even ration of live biomass (shrimp and microbial flocs) and the lack water
in T4, where the pH was below the recommended range for the best exchange, when increased alkalinity and levels pH reduction the CO2
performance of nitrifying bacteria (7–9; Chen et al., 2006), we did not concentrations increase. Here we found a higher accumulation of dis-
measure disequilibrium in the NO2–N–NO3–N oxidation. According to solved CO2 in T4, which may have affected the shrimp development,
Ganguly et al. (1999), the use of 0.1–0.2 g/L of calcium hydroxide is because CO2 concentrations remained within the harmful range for
 Author's personal copy
134 P.S. Furtado et al. / Aquaculture 321 (2011) 130–135
the species (Van Wyk and Scarpa, 1999). In T3, in which the pH was
close to neutrality but alkalinity was high, the values of CO2 increased
and were within the acceptable range for the species (Van Wyk and
Scarpa, 1999). T1 and T2 showed the lowest CO2 concentrations and,
their zootechnical performance was probably not negatively affected.
According to Van Wyk and Scarpa (1999), L. vannamei need alkalinity
values higher than 100 mg CaCO3/L for good development. However,
Ebeling et al. (2006) found that in systems with limited water ex-
change, the alkalinity should be between 100 and 150 mg CaCO3/L.
In this study, the alkalinity concentrations decreased in all treat-
ments. In T4 it decreased from 160 to 35 mg CaCO3/L in 60 days,
whereas in T1 (in which no correction was performed either), the
minimum value of 55 mg CaCO3/L was observed. In this treatment
the alkalinity concentrations were higher than in T4 because the car-
bonate material used to increase the pH has increased the alkalinity
by approximately 20 mg CaCO3/L.
These data corroborate those obtained by McIntosh (2001), which
affirm that the values of alkalinity can decrease to less than 20 mg
CaCO3/L if carbonate material is not used. For Whangchai et al.
(2004), the calcium carbonate was not efficient in increasing the alka-
linity, but high purity sodium bicarbonate and calcium hydroxide
were efficient in controlling the alkalinity of a shrimp culture treated
with ozone. The correction of alkalinity through the use of sodium bi-
carbonate and calcium hydroxide, in contrast to sodium carbonate,
was efficient in increasing the alkalinity in BFT cultures.
Marine shrimp present their best development in cultures with pH
range from 7.0 to 9.0 (Van Wyk and Scarpa, 1999). The reduction of
the alkalinity concentration in T4 negatively affected the pH values,
which remained below the ideal levels for culture for the last 28 exper-
imental days (Van Wyk and Scarpa, 1999). This also negatively affected
the shrimp performance. In T1, the alkalinity concentrations decreased
below the adequate range for the species (Ebeling et al., 2006). Howev-
er, it did not negatively affect the pH levels because they were corrected
with sodium carbonate and, thus, remained within the ideal range for
the species (Van Wyk and Scarpa, 1999). Thus, the efficiency of sodium
carbonate in increasing the pH was verified. The T3 and T2 did not show
alkalinity levels below the adequate range for the good development of
the species (Van Wyk and Scarpa, 1999); however, the pH concentra-
tions in T3 showed values below 7 in the last eight experimental days,
whereas the pH levels in T2 remained within the ideal range for best
growth of the juveniles. Typically in heterotrophic systems, pH de-
creases during the culture. This was clearly demonstrated in T4. Vinatea
et al. (2010) observed a reduction in pH correlating with the increase in
respiration in BFT systems. This corroborates the data obtained in this
study and the studies performed by Wasielesky et al. (2006b). Survival
rates were similar results were obtained by Ray et al. (2010), working
with L. vannamei juveniles (460 shrimp/m 3), obtained survival rates of
71± 8%.
For weight gain and final weight, the best results were obtained in
T1. T2 and T3 were not significantly different, but they were greater
than in T4. In T4, there was unsatisfactory growth at both laboratory
and commercial scales. In relation to the weight gain/week, there were
significant differences (P b 0.05) in T1, T2 and T3 (1.1, 1.0 and 1.01 g, re-
spectively) when compared to T4 (0.73 g). These values are higher than
the 0.65 and 0.89 g obtained, respectively, by Krummenauer (2008)
working with 300 shrimp/m2 in southern Brazil, and Ray et al. (2010)
working with systems of total suspended solids removal and stocking
densities of 460 shrimp/m3; however, they are lower than the results
obtained by Wasielesky et al. (2006a) (300 shrimp/m2) which regis-
tered weight gain/week of 1.25 g with L. vannamei juveniles reared in
BFT system. The feed conversion rate verified in the treatments in
which there were no uses of alkalinizing compounds was similar to
those obtained by Krummenauer (2008) and Otoshi et al. (2007), who
obtained values of 1.27 and 1.49, respectively. However, the feed con-
version rate registered in T4 was approximately two times higher than
those verified by the cited authors. Thus, this value of feed conversion
rate is inadequate for the commercial culture of L. vannamei in BFT
system.
The highest values of final biomass and productivity were ob-
served in T1 and T2, followed by T3 and T4. The values of productivity
in this study were lower than those obtained by Krummenauer
(2008). In the literature, good productivity rates are typically around
7.5 kg/m 3 (Otoshi et al., 2007). However, to obtain these values it is
necessary to use additional technological methods, such as injection
of liquid oxygen, the use of probiotics, artificial substrates, external
filters for total suspended solids removal and control, balanced diets
specially for BFT systems and/or shrimp genetically selected for great-
er growth potential (speed line).
5. Conclusion
The data obtained in this study demonstrates that calcium hydrox-
ide and sodium bicarbonate were efficient compounds for increasing
the alkalinity in BFT cultures, whereas sodium carbonate was not ef-
fective in increasing the alkalinity. However, sodium carbonate was
efficient in increasing the pH and contributed to the increase of alka-
linity. This clearly shows, through the results obtained from T4 that
the levels of alkalinity and pH decrease, whereas concentrations of
CO2 increase in super-intensive BFT systems with use of alkalinizing
compounds. Additionally, the water quality and the zootechnical per-
formance were negatively affected when the concentrations of alka-
linity remain below 100 mg CaCO3/L and pH 7 for extended periods.
Thus, in BFT culture systems, it is necessary to increase the levels of
alkalinity and pH through the use of alkalinizing compounds.
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