Aquaculture Reports 20 (2021) 100709

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Aquaculture Reports
journal homepage: www.elsevier.com/locate/aqrep

Effect of nursery system and stocking density to produce juveniles of

whiteleg shrimp Litopenaeus vannamei
Delia Rodríguez-Olague a, Jesús T. Ponce-Palafox a, b, *, 1,
Sergio G. Castillo-Vargasmachuca a, b, *, 1, Eulalio Arámbul-Muñoz a, Raúl C. de los Santos a,
Héctor M. Esparza-Leal c
a
Maestría en Ciencias Biológicas y Agropecuarias (CBAP), Universidad Autónoma de Nayarit, Mexico
b
Laboratorio de Bioingeniería Costera. Escuela Nacional de Ingeniería Pesquera. Universidad Autónoma de Nayarit, Mexico
c
Departamento de Acuacultura, Instituto Politécnico Nacional-CIIDIR Unidad Sinaloa, Guasave, Sinaloa, Mexico

A R T I C L E I N F O A B S T R A C T

Keywords: Indoor nursery systems are becoming increasingly popular for high-quality whiteleg shrimp production. These
Clean-water systems allow for a significant degree of control and management in a biosecure manner. However, it is still
Photo-heterotrophic unclear as to what type of production system is best suited for indoor whiteleg shrimp nurseries. This study
Biofloc technology
evaluated and compared clean-water recirculating aquaculture system (CW-RAS), photo-heterotrophic (PT), and
Carrying capacity
Indoor tanks nursery
biofloc technology (BFT) systems in terms of water quality, growth performance, and production costs of
whiteleg shrimp Litopenaeus vannamei at three stocking densities (500, 1000 and 1500 org/m3) for 35 d in the
nursery phase. In this study, twenty-seven, 100-L circular plastic tanks (microcosm) were randomly assigned to
one of the treatments and stored with postlarvae shrimps (0.016 g). Throughout the experiment, water quality
parameters remained within acceptable ranges for whiteleg shrimp growth. High concentrations of total
ammonia-nitrogen (TAN) were determined in the CW-RAS system and nitrates and nitrites in the BFT system.
Furthermore, the growth parameters showed significant differences (P < 0.05) between the treatments; higher
values in PT and BFT systems at 500 org/m3 and lower survival in the CW-RAS system. These results suggest that
a dense-dependent effect was observed from 1000 org/m3 onwards. In general, postlarvae, feed, labor and energy
accounted for higher costs. The study findings further indicate that CW-RAS had the highest production costs for
1000 juveniles.

1. Introduction from the original model Generation 1 (large ponds and low production)
to the current Generation 5 (small ponds, total water recirculation, and
The culture of whiteleg shrimp, Litopenaeus vannamei continues to waste capture with more environmental controls for high production),
expand worldwide, reaching a production of 4,966,200 MT (FAO, which signifies a model of high biosecurity (McIntosh, 2019). The latter
2020). Over the past three years, the production increased at a com­ model generally includes controlled systems with smaller ponds and
pound annual growth rate (CAGR) of 5.7 percent (Anderson et al., tanks, shaded, and covered to reduce environmental fluctuations.
2019). However, in recent times, environmental degradation, climate Additionally, it requires healthy postlarvae free from diseases and
change, and the proliferation of pathogens have emerged as key chal­ modern hatcheries with sanitation, quality control, automation, recy­
lenges that are preventing the sustainable development of the world’s cling of water, and reduction in the use of fresh food (Jory, 2019).
whiteleg shrimp farming industry (Jory, 2019). Additionally, interna­ As with other industries, productivity remains a key factor affecting
tional market prices, feed, fuel costs, and whiteleg shrimp diseases the bottom line of whiteleg shrimp producers. Consequently, the two-
continue to have a negative influence on Latin America’s whiteleg phase system that includes a nursery stage followed by the grow-out
shrimp producers (Anderson et al., 2019). stage has been implemented for more than two decades in order to
Over the last five decades, whiteleg shrimp farming has changed improve the quality of whiteleg shrimp production at a super-intensive

* Corresponding authors at: Laboratorio de Bioingeniería Costera. Escuela Nacional de Ingeniería Pesquera. Universidad Autónoma de Nayarit, Mexico.
E-mail addresses: [email protected] (J.T. Ponce-Palafox), [email protected] (S.G. Castillo-Vargasmachuca).
1
These authors contributed equally to this work.

https://doi.org/10.1016/j.aqrep.2021.100709
Received 11 August 2020; Received in revised form 20 April 2021; Accepted 20 April 2021
Available online 30 April 2021
2352-5134/© 2021 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
D. Rodríguez-Olague et al. Aquaculture Reports 20 (2021) 100709

level (Samocha and Lawrence, 1998). In whiteleg shrimp production An analysis of the effect of stocking density of L. vannamei in the BFT
intensification, the density of the shrimp population remains the most system has been undertaken comparing with CW-RAS system as a
important parameter. Several physico-chemical variables and popula­ controlled environment. Most studies suggest that no differences exist
tion parameters, such as specific growth rate (SGR) and survival rate, between the two systems with a non-significant trend for better growth
tend to be critical at extreme limits, in high densities. On the basis of (Esparza-Leal et al., 2015; Ray et al., 2017; Martinez-Porchas et al.,
these parameters, the most appropriate density of the whiteleg shrimp 2020). The implementation and selection of these RAS for the produc­
L. vannamei population in the grow-out phase in the biofloc technology tion of whiteleg shrimp L. vannamei in the nursery phase remain reliant
(BFT) system is 500–625 org/m2 (Da Silveira et al., 2020). However, it is on the location. However, it is also essential to consider the availability
imperative to note that the quality of postlarvae management in the of inputs and production strategies as well as the understanding of
nursery phase is a key factor for their growth and survival as well as the physical-chemical, biological, and economic factors for
basis for juveniles’ maximum growth performance in the grow-out phase decision-making. (Juarez and Martinez-Cordero, 2004). Thus, the study
(Zelaya et al., 2007). of production costs from the nursery phase, along with other factors,
In the whiteleg shrimp farming industry, the nursery systems provide helps deepen the understanding of the whiteleg shrimp industry’s
numerous benefits, including improved inventory, increase in size uni­ overall economy. The operation costs particularly depend on services,
formity, less predation, reduced pond grow-out duration, an increase in supplies, size of the operation, design of the nursery system (Naegel,
the number of harvests per year, better feed conversion efficiencies, and 2010), and energy expenditure. Accordingly, a bioeconomic analysis
greater biosecurity (Persyn and Aungst, 2001; Arnold et al., 2006). Over and stochastic model have been performed to elucidate the single-phase
the past two decades, a series of strategies have been used to intensify production dynamics of a photo-heterotrophic system (Moreno-Figueroa
the nursery phase of whiteleg shrimp production. This includes the use et al., 2019), where feed and postlarvae account for the main costs. Thus
of high-protein feeds, fertilizers, indoor recirculating raceways, and far, no studies have been carried out in the nursery phase. The com­
artificial substrates (Moss and Moss, 2004). Consequently, in the middle parison of different RAS is required to include the elements of decision
of the last decade, recirculating aquaculture systems (RAS) technology for their best application. This study aimed to compare three nursery
for nursery whiteleg shrimp production has emerged as the most systems for whiteleg shrimp production – CW-RAS, PT, and BFT in terms
advanced and successful system with high survival (52.0%–97.5%) and of water quality, growth performance, survival, and production costs at
high densities of 3,000–8,300 org/m3 (Davis and Arnold, 1998; Cohen different stocking densities.
et al., 2005; Mishra et al., 2008; Tierney et al., 2020).
Currently, new farming techniques and systems are required to 2. Materials and methods
maximize production in the nursery and grow-out phases, mitigate the
negative effects of heightened stocking density, and generate high 2.1. Location and animals
quality (premium price) whiteleg shrimp at the lowest cost. This has
resulted in the development of improved whiteleg shrimp farming sys­ The experiments were performed in the Coastal Bioengineering
tems in intensive and hyper-intensive controlled systems including CW- Laboratory of the National School of Fishery Engineering, University of
RAS, PT, and BFT systems. Today, these technologies have been suc­ Nayarit, San Blas Nayarit, Mexico (21◦ 29′ N, 105◦ 12′ W). The postlarvae
cessful on a commercial level and can be applied depending on the re­ of L. vannamei were supplied by a commercial laboratory (Acopio de
sources, conditions, and market strategies. Although the CW-RAS Larvas y Asesoría en Proyectos S.A. de C.V.®) located in Matanchen Bay,
maintains high water quality and density, the system can have higher San Blas, Nayarit. Mexico.
start -up and operating costs (Table1). PT system allows for high growth
at low production costs. Furthermore, whiteleg shrimps show good 2.2. System and experimental design
growth and survival in the BFT system, which, entails higher costs than
the PT system. A completely randomized design method with two independent
The PT system is based on the balance between photoautotrophic variables was used in this study. These variables include treatment
(phytoplankton) and heterotrophic (bacteria) organisms (McIntosh, systems (CW-RAS, PT, and BFT) and stocking densities (500 (D500),
2001; Huda et al., 2013; Moreno-Figueroa et al., 2018). In PT and BFT 1000 (D1000) and 1500 (D1500) org/m3). Nine treatments were tested
systems, the density is determined in the nursery and grow-out phases. in triplicate for 35 d. The study was conducted in twenty-seven circular
In the nursery phase, low densities of 1500 org/m3 have been used to indoor polyethylene nursery tanks (0.1 m3) stocked with 28-d postlarvae
high densities of 9000 org/m3, finding the best specific growth rate L. vannamei (initial mean weight of 0.016 g ± 0.002 g) that were hand-
(SGR) at 1500 org/m3 and demonstrating that growth in the nursery counted into the tanks.
phase is density-dependent (Esparza-Leal et al., 2015). In addition, when Water oxygenation in each tank was provided with 1 air diffuser
a substrate is integrated into the BFT, the lateral area of the tank in­ stone and generated by an integrated system of air of 4.5 HP regenera­
creases, which, in turn, causes a higher growth of biofilms and natural tive blower. Then, the aeration was conducted through a plastic hose (2′′
feeds. Consequently, it improves water quality, growth, feed conversion diameter) to the end in each tank. For all systems, the water was sup­
ratio (FCR), and immune parameters of whiteleg shrimp culture (Fer­ plied directly from the sea through PVC pipe (3′′ diameter) with a
reira et al., 2015; Kumar et al., 2019). Jacuzzi Magnum force3® type pump, which passed through two Life­
gard® Aquatic’s filters with zeolite sand and activated carbon, respec­
Table 1 tively also ending in the culture units. Both aeration and water lines
Factor and level of the different types of systems studied in the culture of white were disinfected with 5% formalin and 30 % hydrochloric acid,
shrimp L. vannamei. Clear-water recirculating aquaculture systems (CW-RAS), respectively. There was no renewal of water in all systems during the
photo-heterotrophic (PT), and biofloc technology (BFT). study period; only the water lost due to evaporation was replaced. Each
Factors RAS PT BFT CW-RAS tank had an external biofilter composed of an 8-cm air diffuser;
the water passed through cotton wadding, zeolite sand, and activated
Water quality
carbon, thus leaving a drain at the bottom and returning to the shrimp
+++ ++ +
Productivity natural – + ++
Density +++ ++ + tank through a pump "Power Head SPH-800.′′ An internal support tube
Specific growth rate + +++ ++ maintained the water level, ensuring the exchange of water between a
Survival ++ +++ ++ filter and the culture tank.
Costs +++ + ++
Cyclotella choctawhatcheeana microalgae (180,000 cel/mL) was
Importance level factor: +, low; ++, medium; +++, high; -, nil. added to the PT system on days 1, 5, 10, and 15 of the experiment with a

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D. Rodríguez-Olague et al. Aquaculture Reports 20 (2021) 100709

proportion equivalent to 8% of the total volume of each tank to maintain 2.6. Data management and analysis
a C:N ratio of 2–3 (Martinez-Porchas et al., 2020). The microalgae in­
oculums were obtained from Commercial Production Laboratory “Aco­ The normality tests (Shapiro-Wilk) and homoscedasticity (Levene)
pio de Larvas y Asesoría en Proyectos S.A. de C.V®”. In addition, a were determined for each data group. Additionally, the variables were
commercial probiotic (Neutrobacter®) was added to each tank once per analyzed using the One-way Analysis of Variance (ANOVA) and post-hoc
week until the end of the trial (250 mL/m3) with a view to promoting a Tukey test (Zar, 2010). In all cases, the level of significance was found to
heterotrophic microbial complex. Furthermore, 10 % of molasses per kg be 5% (P < 0.05). Before the analysis, the survival data were trans­
of feed was added to each tank at the beginning of the experiment so as formed (arcsine of the square root) and expressed as mean ± standard
to improve nitrification. deviation. Using the data obtained in the study, the projections of the
In the BFT system, biofloc water in the experimental tanks came from density-production relationship were made up to a density of 9000
a matured biofloc shrimp tank (with 0.935 mg/L, 0.325 mg/L, 3.392 org/m3. This density was obtained under the similar conditions of pre­
mg/L, 0.564 mg/L, and 180.0 mg/L of total ammonia nitrogen, nitrite, vious works comparing CW-RAS with BFT in L vannamei (Esparza et al.,
nitrate, phosphate, and total suspended solids, respectively), where 2015). The extrapolation of the density-production relationship was
molasses, wheat flour, balanced feed, and shrimp were previously performed in accordance with the criteria of López-Uriostegui et al.
applied. The C:N ratio for BFT was adjusted to 12:1 by adding molasses (2014), where the relationship produced a quadratic equation in the
as a carbon source weekly and putting plastic mesh in each tank as a form:
substrate to generate the formation and proliferation of biofilm and
biofloc (Marinho et al., 2017). Production = β + β + β2d2, (1)
The postlarvae were fed a commercial feed containing 0.9–1.1 mm where β2 is a quadratic coefficient (other than 0), β1 signifies the linear
40 % crude protein (Maltacleyton®, Mexico) at different times of day coefficient, β0 is the intercept, and d denotes the density
(0800, 1400, and 2000 h). The daily feeding rates were initially at The maximum density (dmax) was obtained as follows:
16− 10% of estimated biomass. Subsequently, the feed was adjusted
daily according to its consumption, to determine the consumption; 10 % dmax = β1/ 2β2, (2)
of the pellet was placed in trays and the rest was distributed in the tank.
where dmax = maximum density.
The maximum production was given by

2.3. Water analysis Productionmax = β0 + ((β1)2 / 2β2) - β1/2, (3)

The experimental area’s photoperiod was 12/12 h light/dark cycle.

The water temperature was maintained by two Minisplit Mabe Cold and
Heat 12,000 BTUs placed in strategic areas that regulated the temper­ 3. Results
ature of the experimental room. For each tank, the dissolved oxygen (DO
mg/L) was recorded on a daily basis every 6 h (8:00, 14:00, 20:00, and 3.1. Water physical and chemical variables
02:00 h) with YSI 550A® (YSI Incorporated, Yellow Springs, Ohio, USA).
Additionally, the salinity and pH were recorded twice a day (7:00 and The variables of temperature, salinity, pH, and OD did not show
19:00) with a Vital Sine SR-6® refractometer and Hanna pHep Tester® significant differences (P > 0.05) in all treatments throughout the 35 d of
potentiometer, respectively. Every third day, the YSI 9500 direct-read the experiment. The mean values and standard deviation (SD) for these
photometer® (YSI Incorporated, Yellow Springs, Ohio, USA) was used variables are shown in Table 2. Among all the tested densities, the
to measure total ammonia nitrogen (TAN), non-ionized ammonium highest concentrations of TAN were observed in the CW-RAS treatment,
(NH3-N), nitrite (NO2-N), and nitrate (NO3-N) concentrations in mg/L. reaching a mean concentration of 1.23 mg/L in D1500. On the other
hand, the highest concentrations of NO2-N and NO3-N were determined
in the BFT treatment in all the tested densities, reaching a maximum
2.4. Growth and production performance concentration from 3.45 to 3.92 mg/L and 10.65–13.48 mg/L,
respectively.
Biometrics was performed at the beginning and end of the experi­
ment by weighing, measuring, and counting all the shrimps from each 3.2. Growth and production performance
experimental tank using a digital balance (precision 0.001 g, Sartorius®)
and a stainless steel metal ruler with conversion table 30.5 ± 0.5 cm. In general, differences (P < 0.05) were found in growth performance,
Initial body weight (IBW, g) and initial biomass (IB, g) were determined use of feed, and production parameters in all the treatment systems and
at the beginning of the study. After 35 d, at the end of the study, the total densities. The whiteleg shrimps grew significantly larger with greater
number of shrimp (TNS), final body weight (FBW, g), final length (FL, survival in the PT and BFT treatments than in CW-RAS (Table 3). It
cm), condition factor (CF), total feed consumption (TFC, g), feed con­ caused higher biomass and production in the density of 1500 org/m3
version factor (FCR), feed efficiency (FE), final biomass (BF, g), Pro­ with a better feed conversion ratio and feed efficiency in the BFT
duction (P, kg/m3), variation coefficient-body weight (CV-BW, %), treatment. However, the coefficient of variation of the whiteleg shrimp
weight gain (WG), growth rate (GR, g/week), specific growth rate (SGR, was observed to be lower in the BFT treatment in all densities.
%/d), and survival (S, %) were determined. The effect of density on the whiteleg shrimp production in CW-RAS,
PT, and BFT systems was described by a quadratic (or parabolic) func­
tion – a polynomial function of the second degree described in Fig. 1.
2.5. Production costs Where the equation for CW-RAS, BF and PT were y = -2E-08x2 +
0.0003x + 0.0625, R2 = 0.9978; y = -2E-08x2 + 0.0004x + 0.3432, R2 =
All the production costs of each treatment were recorded. Expendi­ 0.998 and y = -2E-08x2 + 0.0004x + 0.5688, R2 = 0.9911, respectively.
tures were shown as costs per whiteleg shrimp produced per cubic meter The carrying capacity (CC) at 8000 org/m3 for PT and BFT and 9000
and cost per 1000 juvenile of whiteleg shrimp produced at the end. The org/m3 for CW-RAS (Fig. 1a)
production costs were grouped into 8 categories: postlarvae, feed,
microalgae, probiotics, chemicals, operating supplies, labor, and energy
(Juarez and Martinez-Cordero, 2004; Naegel, 2010).

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Table 2
Physical and chemical variables of water of the L. vannamei in clear-water recirculating aquaculture systems (CW-RAS), photo-heterotrophic (PT), and biofloc
technology (BFT) treatments during the nursery phase cultured indoor at 500 (D500), 1000 (D1000) and 1500 (D1500) orgs/m3 over 35-d.

Parameters/ D500 D1000 D1500

treatment CW-RAS PT BFT CW-RAS PT BFT CW-RAS PT BFT

Temperature (ºC) 29.5 ± 0.7a 29.4 ± 0.6a 29.4 ± 0.7a 29.3 ± 0.7a 29.4 ± 0.6a 29.4 ± 0.7a 29.4 ± . 0.8a 29.4 ± . 0.1a 29.4 ± . 0.6a
Salinity 34.9 ± 0.1a 34.9 ± 0.1a 34.9 ± 0.1a 34.9 ± 0.1a 34.9 ± 0.1a 34.9 ± 0.1a 34.9 ± 0.1a 34.9 ± 0.1a 34.9 ± 0.1a
pH 8.47 ± 0.07a 8.42 ± 0.11a 8.45 ± 0.11a 8.42 ± 0.13a 8.39 ± 0.12a 8.45 ± 0.11a 8.44 ± 0.06a 8.41 ± 0.11a 8.41 ± .09a
Oxygen (mg/L) 6.24 ± 0.75a 6.35 ± 0.72a 6.48 ± 0.69a 6.14 ± 0.75a 6.25 ± 0.72a 6.37 ± 0.69a 6.11 ± 0.70a 6.17 ± 0.76a 6.25 ± 0.69a
TAN (mg/L) 1.11 ± 1.17a 0.49 ± 0.29b 0.93 ± 0.72a 1.19 ± 1.28a 0.51 ± 0.34b 1.06 ± 0.85a 1.23 ± 1.17a 0.65 ± 0.46b 1.09 ± 0.93a
NO2-N (mg/L) 0.34 ± 0.26c 2.20 ± 1.15b 3.69 ± 1.18a 0.40 ± 1.26c 2.08 ± 1.16b 3.42 ± 1.11a 0.67 ± 0.31c 2.11 ± 1.18b 3.95 ± 1.12a
NO3-N (mg/L) 4.29 ± 2.12b 1.50 ± 1.02c 10.65 ± 2.30a 5.61 ± 2.98b 1.56 ± 0.89c 11.38 ± 2.67a 9.5 ± 2.16b 1.69 ± 0.85c 13.48 ± 2.35a

Within a row, values (mean ± SD) accompanied by different letters are significantly different (P < 0.05). TAN, total ammonia nitrogen; NO2-N, nitrite; and NO3-N,
nitrate.

Table 3
Growth performance, survival and distribution size of the L. vannamei in clear-water recirculating aquaculture systems (CW-RAS), photo-heterotrophic (PT), and
biofloc technology (BFT) treatments, during the nursery phase cultured indoor at 500 (D500), 1000 (D1000) and 1500 (D1500) orgs/m3 over 35-d.

Parameters/ D500 D1000 D1500

treatment CW-RAS PT BFT CW-RAS PT BFT CW-RAS PT BFT

IBW (g/org) 0.016 ± 0.016 ± 0.016 ± 0.016 ± 0.016 ± 0.016 ± 0.016 ± 0.016 ± 0.016 ±
0.007a 0.007a 0.007a 0.007a 0.007a 0.007a 0.007a 0.007a 0.007a
FBW (g/org) 0.44 ± 0.05d 1.27 ± 0.09a 1.21 ± 0.03a 0.42 ± 0.06d 0.95 ± 0.09b 0.91 ± 0.02b 0.41 ± 0.05d 0.82 ± 0.09c 0.80 ± 0.03c
FL (cm) 4.59 ± 0.11c 5.42 ± 0.50a 6.02 ± 0.36a 4.38 ± 0.51c 5.35 ± 0.56a 5.86 ± 0.16a 4.57 ± 0.33c 5.20 ± 0.57a 5.38 ± 0.18a
CF (%) 0.46 ± 0.03d 0.80 ± 0.12a 0.55 ± 0.06b 0.45 ± 0.16d 0.62 ± 0.14b 0.45 ± 0.04d 0.43 ± 0.05d 0.58 ± 0.11b 0.50 ± 0.02c
FB (g) 203.0 ± 3.2d 655.1 ± 5.5b 550.2 ± 4.9b 347.1 ± 7.2c 915.3 ± 7.7a 693.1 ± 5.8b 493.2 ± 1150.2 ± 9.6a 856.9 ± 9.8a
9.62c
P (kg/m3) 0.20 ± 0.00d 0.65 ± 0.00b 0.55 ± 0.00b 0.35 ± 0.00c 0.92 ± 0.00a 0.69 ± 0.00b 0.49 ± 0.01c 1.15 ± 0.01a 0.86 ± 0.01a
TFC (g) 21.53 ± 67.10 ± 53.87 ± 47.18 ± 92.17 ± 84.16 ± 68.18 ± 125.20 ± 118.82 ±
2.11e 11.12c 5.99c 6.23d 29.07b 10.05b 5.37c 29.83a 13.73a
FCR 1.39 ± 0.27b 1.34 ± 0.12b 1.21 ± 0.14a 1.78 ± 0.31d 1.46 ± 0.24c 1.28 ± 0.14a 1.82 ± 0.29d 1.48 ± 0.23c 1.40 ± 0.21c
FE (%) 0.72 ± 0.02b 0.72 ± 0.01b 0.83 ± 0.01a 0.56 ± 0.03c 0.68 ± 0.02b 0.78 ± 0.02a 0.55 ± 0.02c 0.68 ± 0.03b 0.71 ± 0.02b
CV-BW 9.09 ± 0.03c 7.12 ± 0.02b 2.50 ± 0.01a 13.11 ± 10.36 ± 2.64 ± 0.03a 11.44 ± 9.10 ± 0.02c 3.44 ± 0.02a
0.02c 0.01c 0.04c
WG (g/org) 0.42 ± 0.02d 1.25 ± 0.07a 1.19 ± 0.03a 0.40 ± 0.05d 0.93 ± 0.04b 0.89 ± 0.03b 0.39 ± 0.02d 0.80 ± 0.04c 0.78 ± 0.03c
GR (g/week) 0.012 ± 0.036 ± 0.034 ± 0.012 ± 0.027 ± 0.026 ± 0.011 ± 0.023 ± 0.022 ±
0.001c 0.003a 0.002a 0.001c 0.002b 0.002b 0.001c 0.003b 0.003b
SGR (%/d) 9.47 ± 0.03c 12.50 ± 12.36 ± 9.34 ± 0.33c 11.67 ± 11.55 ± 9.27 ± 0.22c 11.25 ± 0.02b 11.18 ± 0.15b
0.44a 0.36a 0.15b 0.07b
S (%) 92.1 ± 2.5b 98.9 ± 4.1a 98.5 ± 2.5a 82.5 ± 3.5b 96.3 ± 1.9a 95.2 ± 3.6a 80.1 ± 4.1b 94.6 ± 4.3a 93.3 ± 2.6a

Within a row, values (mean ± SD) accompanied by different letters are significantly different (P < 0.05). IBW, initial body weight; FBW, final body weight; FL, final
length; CF, condition factor; FB, final biomass; P, production; TFC, total feed consumed; FCR, feeding conversion ratio; FE, feeding efficiency; CV-BW, coefficient of
variation- body weight; WG: weight gain; GR: growth rate; and SGR: specific growth rate; S, survival.

3.3. Production costs

Four types of costs contributed over 85 % of the total production

costs in all treatments (Table 4). The highest costs were in the order of
importance – labor, postlarvae, energy, and feed – for CW-RAS. The
order of importance was labor, postlarvae, feed, and microalgae for PT
and labor, postlarvae, chemical, and feed for BFT.

4. Discussion

Temperature, salinity, DO, and pH remained within acceptable

ranges in all treatments for the growth of L. vannamei (Ponce-Palafox
et al., 2019). The levels of TAN, nitrites, and nitrates in all treatments
were within the range recommended for whiteleg shrimp culture
(Valencia-Castañeda et al., 2019). However, the TAN was higher in the
CW-RAS treatment as compared to the PT and BFT treatments. Addi­
tionally, all the measured TAN concentrations were similar or below the
Fig. 1. Relationship between production (color lines) and survival (black lines) estimated safe levels for rearing L. vannamei in salinities near 35 (Lin and
of whiteleg shrimp L. vannamei at different densities in nursery system. Carrying Chen, 2001). In the PT and BFT treatments, the lowest concentration of
capacity (a). The curves of the treatments of CW-RAS (circle), BFT (square) and TAN showed that the three TAN elimination processes described in
PT (triangle) are shown. aquatic systems were presented in these treatments: photoautotrophic
elimination by algae, autotrophic bacterial conversion, and heterotro­
phic bacterial conversion directly into microbial biomass (Ebeling et al.,
2006). The maximum mean nitrite concentration recorded in all

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Table 4
Comparative analysis of production costs of the L. vannamei in clear-water recirculating aquaculture systems (CW-RAS), photo-heterotrophic (PT), and biofloc
technology (BFT). Treatments system during the nursery phase cultured indoor at 500 (D500), 1000 (D1000) and 1500 (D1500) orgs/m3 over 35-d.
D500 D1000 D1500
Parameters/
treatment CW- % PT % BFT % CW- % PT % BFT % CW- % PT % BFT %
RAS RAS RAS

Postlarvae 2.25 30.0 2.25 37.4 2.25 31.9 4.5 42.0 4.5 51.3 4.5 45.6 6.8 49.2 6.8 57.9 6.8 52.9
Feed 0.59 7.9 0.58 9.6 0.49 6.9 1.54 14.4 1.09 12.4 1.05 10.6 2.35 17.0 1.76 15.0 1.73 13.5
Microalgae – 0.0 0.44 7.3 – 0.0 – 0.0 0.44 5.0 – 0.0 – 0.0 0.44 3.7 – 0.0
Probiotic – 0.0 0.19 3.2 – 0.0 – 0.0 0.19 2.2 – 0.0 – 0.0 0.19 1.6 – 0.0
Chemical 0.14 1.9 0.14 2.3 1.9 26.9 0.14 1.3 0.14 1.6 1.9 19.3 0.14 1.0 0.14 1.2 1.9 14.8
Operating 0.33 4.4 – 0.0 – 0.0 0.33 3.1 – 0.0 – 0.0 0.33 2.4 – 0.0 – 0.0
supplies
Labor 2.3 30.6 2.3 38.2 2.3 32.6 2.3 21.5 2.3 26.2 2.3 23.3 2.3 16.6 2.3 19.6 2.3 17.9
Energy 1.9 25.3 0.12 2.0 0.12 1.7 1.9 17.7 0.12 1.4 0.12 1.2 1.9 13.7 0.12 1.0 0.12 0.9
Total costs ($/m3) 7.51 6.02 7.06 10.71 8.78 9.87 13.82 11.75 12.85
Cost/1000 16.3 12.2 14.3 13.0 9.1 10.4 11.5 8.3 9.2
juvenile ($)

$ = US Dollar.

treatments was lower than the maximum value calculated (25.7 mg/L) (Zelaya et al., 2007). In the nursery phase, this parameter can have a
for L. vannamei (Lin and Chen, 2003). In their study, Ray et al. (2017) wide distribution of size by weight up to 24.0%–70.0% of CV-BW (Moss
found that the nitrite concentrations in CW-RAS were lower than in BFT, and Moss, 2004; Zelaya et al., 2007). However, in our study, CV of the
as recorded in our experiment. This could be due to significantly (P < weight size distribution decreased in the BFT system, which was in
0.05) high rates of water renewal in CW-RAS as compared to other agreement with the findings of other studies in the nursery phase using
systems (Silva et al., 2015). Nitrate, the final product of nitrification, the BFT system (Khanjani et al., 2020). In our study, the CV-BW was low
was observed to have the highest concentration (p < 0.05) in the (2.5 a 13.11 %) and exhibited the pattern: CW-RAS > PT > BFT. This
CW-RAS and BFT treatments (Camargo et al., 2005). The nitrate levels indicates that more than 97.5 % of the whiteleg shrimps in 35 d had a
recorded for all treatments meanwhile, were below the critical levels for relatively uniform weight in the BFT system. Additionally, survival was
L. vannamei (Furtado et al., 2015; Satanwat et al., 2020). Furthermore, not found to be affected by stocking density, as reported in similar
the final nitrate concentration was higher in the BFT system than in studies of the three systems evaluated at different stocking densities
CW-RAS and the pattern was similar to that reported in nursery tanks (Wasielesky et al., 2013; Esparza-Leal et al., 2015;Guemez-Sorhouet
employed in other studies for L. vannamei (Esparza-Leal et al., 2015, et al., 2019; Tierney et al., 2020).
2020; Ray et al., 2017). In the PT system, microalgae have helped in The relationship between production and increased density in an
reducing the concentration of nitrates (Jiménez-Ordaz et al., 2021). intensive RAS of whiteleg shrimp L. vannamei has been established as
However, the heterotrophic pathway does not generate nitrate (Rajta dense-dependent type related through a quadratic function (Williams
et al., 2020), thus justifying the lower concentrations recorded in this et al., 1996). These results suggest that a dense-dependent effect was
system. observed from 8000 to 9000 org/m3 onwards. This effect was critical
The growth and survival parameters in CW-RAS, PT, and BFT sys­ when reaching carrying capacity (Fig. 1, letter "a"). The production
tems were within the range of values considered adequate for the continues to increase, albeit at a decreasing rate (Hepher, 1978). In
nursery phase in intensive whiteleg shrimp farming (Godoy et al., 2012; contrast, survival had a density-independent effect in all the systems
Esparza-Leal et al., 2015; Tierney et al., 2020). The values recorded in evaluated in this study.
this study were higher than those recorded in most experiments of these In the selection of CW-RAS, PT, and FBT systems at a commercial
culture systems. Better growth and productive performance (24.2 %) level, it is recommended to not only consider productive efficiency as a
than the CW-RAS system were observed in the PT and BFT systems, as deciding factor in the nursery phase, but also analyze management
reported in other studies (Izquierdo et al., 2006; Luis-Villaseñor et al., strategies through economic efficiency and optimize the inputs with a
2015; Guemez-Sorhouet et al., 2019). In the nursery phase, we found an view to obtaining the best return on investment (Davis et al., 2008).
inversely proportional relationship between the growth performance of Postlarvae shrimp prices and an increase in variable costs have caused
L. vannamei and stocking density (Wasielesky et al., 2013; Silva et al., stress in the industry. Therefore, the production costs need to be opti­
2015). Additionally, in the present study, FBW, WG, GR, and SGR values mized in principle in order to improve economic returns. In this study,
were higher in the density of 500 org/m3. Growth reduction with the cost structure of the nursing phase of the CW-RAS, PT, and BFT
increased density in the nursery phase has been attributed to decreased systems was closer to that of a hatchery than the grow-out phase, in
water quality, natural productivity, and space (Peterson and Griffith, which the proportion was higher for the costs of labor, postlarvae,
1999; Wasielesky et al., 2013; Silva et al., 2015). As previously reported feeding, and energy (Juarez and Martinez-Cordero, 2004). In addition,
in other studies, the lower FCR value of the PT and BFT systems was the production costs were the highest for the CW-RAS system and the
attributed to the availability of microalgae and microorganisms associ­ lowest for the PT system. In the CW-RAS and BFT systems, we found high
ated with the bioflocs compared to the CW-RAS system (Marinho et al., production and survival of up to 8,000–9,000 org/m3 (Davis and Arnold,
2017). In the current study, the highest feed efficiency was observed in 1998; Cottingham, 2015;Esparza-Leal et al., 2015).
the BFT system at 500− 1,000 org/m3. Moreover, microalgae stocking in
the PT and BFT systems in this study should have contributed to a better 5. Conclusion
performance of the whiteleg shrimp (Godoy et al., 2012; Jiménez-Ordaz
et al., 2021). In this study, the physico-chemical variables of water, temperature,
In the nursery phase, the variability in whiteleg shrimp size was salinity, pH, and DO did not show significant differences in all treat­
caused by size-dependent factors that modify growth and the level of ments. The CW-RAS system had significantly higher levels of TAN, and
overcrowding that can further modify size heterogeneity (Peacor et al., nitrite-nitrate concentrations were found to be higher in BFT. A non-
2007; Araneda et al., 2013). Size uniformity remains a critical aspect in significant tendency was observed to present higher concentrations of
the final harvest of whiteleg shrimp farming at a commercial level TAN and nitrates with increasing density in all treatments. Furthermore,

5
D. Rodríguez-Olague et al. Aquaculture Reports 20 (2021) 100709

growth performance, production, and survival were significantly Esparza-Leal, H.M., Pereira-Cardozo, A., Wasielesky Jr., W., 2015. Performance of
Litopenaeus vannamei postlarvae reared in indoor nursery tanks at high stocking
improved in the PT and BFT treatments in comparison to the CW-RAS
density in clear-water versus biofloc system. Aquac. Eng. 68, 28–34.
treatment. The dense-dependent effect of whiteleg shrimp growth in Esparza-Leal, H.M., Ponce-Palafox, J.T., Álvarez-Ruiz, P., López-Álvarez, E.S., Vázquez-
nursery culture began to critique 7000 org/m3 and 8000 org/m3 in PT- Montoya, N., López-Espinoza, M., Montoya-Mejía, M., Gómez-Peraza, R.L., Nava-
BFT and CW-RAS, respectively. The final results showed that the PT Perez, E., 2020. Effect of stocking density and water exchange on performance and
stress tolerance to low and high salinity by Litopenaeus vannamei postlarvae reared
system had the lowest costs compared to CW-RAS and BFT systems. The with biofloc in intensive nursery phase. Aquac. Int. 28, 1473–1483.
results of this study indicate that the PT system may be a more pro­ FAO, 2020. The State of World Fisheries and Aquaculture 2020. Sustainability in action.
ductive and profitable option than CW-RAR and BFT systems for nursery Rome.
Ferreira, L.M.H., Lara, G., Wasielesky Jr., W., Abreu, P.C., 2015. Biofilm versus biofloc:
whiteleg shrimp production. Finally, it was found that the stocking Are artificial substrates for biofilm production necessary in the BFT system? Aquac.
density in the nursery phase, which registered the best indicators, was Int. 24, 921–930.
500 org/m3. Moreover, the whiteleg shrimp management system and Furtado, P.S., Campos, B.R., Serra, F.P., Klosterhoff, M., Romano, L.A., Wasielesky, W.,
2015. Effects of nitrate toxicity in the Pacific white shrimp, Litopenaeus vannamei,
strategy in the nursery should consider high water quality, growth rate, reared with biofloc technology (BFT). Aquac. Int. 23, 315–327.
and lower costs based on the production goal and conditions in the Godoy, L.C., Odebrecht, C., Ballester, E., Martins, T.G., Wasielesky Jr., W., 2012. Effect of
whiteleg shrimp farm. diatom supplementation during the nursery rearing of Litopenaeus vannamei (Boone,
1931) in a heterotrophic culture system. Aquac. Int. 20, 559–569.
Guemez-Sorhouet, E., Villarreal, H., Racotta, I.S., Naranjo, J., Mercier, L., 2019.
6. Author statement Zootechnical and physiological responses of whiteleg shrimp (Litopenaeus vannamei)
postlarvae reared in bioflocs and subjected to stress conditions during nursery phase.
Aquac. Res. 50, 1198–1211.
Delia Rodríguez Olague: Bioassays, Water Quality, data analysis; Hepher, B., 1978. Ecological aspects of warm water fishpond management. Ch 18. Pp
Jesus T. Ponce-Palafox: Conceptualization, Validation, Reviewing and 447-468. In: Gerking, S.D. (Ed.), Ecology of Freshwater Fish Production. Elsevier
Editing; Sergio G. Castillo-Vargasmachuca: Conceptualization, Vali­ Publishing Co., UK.
Huda, A.S., Ispinanto, J., Bahri, F., Decamp, O., 2013. Successful production in semi-
dation, Reviewing and Editing; Eulalio Arámbul Muñoz; Investigation,
biofloc in Indonesia. Aquac. Asia Pac. 2, 8–12.
Supervision; Raúl Claro de los Santos: Water Quality analysis; Héctor Izquierdo, M., Forster, I., Divakaran, S., Conquest, L., Decamp, O., Tacon, A., 2006. Effect
M. Esparza-Leal: Conceptualization, Validation. of green and clear water and lipid source on survival, growth and biochemical
composition of Pacific white shrimp Litopenaeus vannamei. Aquac. Nut. 12, 192–202.
Jiménez-Ordaz, F., Cadena-Roa, M., Pacheco-Vega, J., Rojas-Contreras, M., Tovar-
7. Ethical approval Ramírez, D., Arce-Amezquita, P., 2021. Microalgae and probiotic bacteria as biofloc
inducers in a hyper-intensive Pacific white shrimp (Penaeus vannamei) culture. Lat.
Am. J. Aquat. Res. 49 (1), 155–168.
All applicable international, national, and/or institutional guidelines
Jory, D.E., 2019. Around the world of shrimp: notes from INFOFISH 2019. Global
for the care and use of animals were followed by the authors. Aquacult. Advoc. (November), 1–6. https://www.aquaculturealliance.org/advocat
e/around-the-world-of-shrimp-notes-from-infofish-2019/?headlessPrint=AAAAA
PIA9c8r7gs82oWZBA.
Declaration of Competing Interest Juarez, L.M., Martinez-Cordero, F.J., 2004. Operating costs and health management
strategies in shrimp hatcheries. Aquac. Econ. Manag. 8, 193–200.
Khanjani, M.H., Alizadeh, M., Sharifinia, M., 2020. Rearing of the Pacific white shrimp,
The authors declare that they have no conflict of interest. Litopenaeus vannamei in a biofloc system: the effects of different food sources and
salinity levels. Aquacult. Nutr. 26, 328–337.
Acknowledgements Kumar, S., Shyne-Anand, P.S., De, D., Ghoshal, T.K., Alavandi, S.V., Vijayan, K.K., 2019.
Integration of substrate in biofloc based system: effects on growth performance,
water quality and immune responses in black tiger shrimp, Penaeus monodon culture.
We want to thank Universidad Autónoma de Nayarit for the financial Aquac. Res. 50, 2986–2999.
support to the project, from which the results of this paper were Lin, Y.C., Chen, J.C., 2001. Acute toxity of ammonia in Litopenaeus vannamei Boone
juveniles at different salinity levels. J. Exp. Mar. Biol. Ecol. 259, 109–119.
obtained.
Lin, Y.C., Chen, J.C., 2003. Acute toxicity of nitrite on Litopenaeus vannamei (Boone)
juveniles at different salinity levels. Aquaculture 224, 193–201.
References López-Uriostegui, F., Ponce-Palafox, J.T., Arredondo-Figueroa, J.L., Benítez-
Mandujano, M.A., García-Ulloa Gómez, M., Castillo-Vargasmachuca, S., Esparza-
Leal, H.M., 2014. Effect of stocking density on growth and survival of the prawn
Anderson, J.L., Valderrama, D., Jory, D.E., 2019. GOAL 2019: global shrimp production
Macrobrachium tenellum cultured in a cage-pond system. N. Am. J. Aquac. 76 (2),
review. Global Aquacult. Advoc (November), 1–5. https://www.aquaculturealliance.
164–169.
org/advocate/goal-2019-global-shrimp-production-review/?headlessPrint=AAAAA
Luis-Villaseñor, E., Voltolina, D., Audelo-Naranjo, J.M., Pacheco-Marges, M.R., Herrera-
PIA9c8r7gs82oWZBA.
Espericueta, V.E., Romero-Beltrán, E., 2015. Effects of biofloc promotion on water
Araneda, M.E., Hernández, J.M., Gasca-Leyva, E., Vela, M.A., 2013. Growth modelling
quality, growth, biomass yield and heterotrophic community in Litopenaeus vannamei
including size heterogeneity: application to the intensive culture of white shrimp
(Boone, 1931) experimental intensive culture. Ital. J. Anim. Sci. 14 (3), 332–337.
(P. vannamei) in freshwater. Aquac. Eng. 56, 1–12.
Marinho, Y.F., Brito, L.O., Silva-Campos, C.V., Severi, W., Andrade, H.A., Galvez, A.O.,
Arnold, S.J., Sellars, M.J., Crocos, P.J., Coman, G.J., 2006. Intensive production of
2017. Effect of the addition of Chaetoceros calcitrans, Navicula sp. And Phaeodactylum
juvenile tiger shrimp Penaeus monodon: an evaluation of stocking density and
tricornutum (diatoms) on phytoplankton composition and growth of Litopenaeus
artificial substrates. Aquaculture 261, 890–896.
vannamei (Boone) postlarvae reared in a biofloc system. Aquac. Res. 48, 4155–4164.
Camargo, J.A., Alonso, A., Salamanca, A., 2005. Nitrate toxicity to aquatic animals: a
Martinez-Porchas, M., Ezquerra-Brauer, M., Mendoza-Cano, F., Higuera, J.E.C., Vargas-
review with new data for freshwater invertebrates. Chemosphere 58, 1255–1267.
Albores, F., Martinez-Cordova, L.R., 2020. Effect of supplementing heterotrophic and
Cohen, J.M., Samocha, T.M., Fox, J.M., Gandy, R.L., Lawrence, A.L., 2005.
photoautotrophic biofloc, on the production response, physiological condition and
Characterization of water quality factors during intensive raceway production of
post-harvest quality of the whiteleg shrimp, Litopenaeus vannamei. Aquacult. Rep. 16,
juvenile Litopenaeus vannamei using limited discharge and biosecure management
100257.
tools. Aquac. Eng. 32, 425–442.
McIntosh, D., 2001. Changing paradigms in shrimp farming: V. Establishment of
Cottingham, G., 2015. An Evaluation of Management Criteria for Biofloc Systems Used in
heterotrophic bacterial communities. Global Aquacult. Advoc. 4 (1), 53–58.
High-density Early Life Stage Rearing of Marine Penaeid Shrimp at Low and High
McIntosh, R., 2019. Modelling shrimp industry towards sustainability. INFOFISH 2019.
Salinity. Thesis degree of Master Science in Fisheries and Mariculture Texas Am
In: World Shrimp Conference and Exposition “Modelling for Sustainability”. 12-14
University – Corpus Christi. Corpus Christi, Texas, USA, 90 pp.
November 2019. Bangkok, Thailand.
Da Silveira, L.G.P., Krummenauer, D., Poersch, L.H., Rosas, V.T., Wasielesky, W., 2020.
Mishra, J.K., Samocha, T.M., Patnaik, S., Speed, M., Gandy, R.L., Ali, A.M., 2008.
Hyperintensive stocking densities for Litopenaeus vannamei grow-out in biofloc
Performance of an intensive nursery system for the pacific white shrimp, Litopenaeus
technology culture system. J. World Aquacult. Soc. 1–11.
vannamei, under limited discharge condition. Aquac. Eng. 38, 2–15.
Davis, D.A., Arnold, C.R., 1998. The design, management and production of a
Moreno-Figueroa, L.D., Naranjo-Paramo, J., Hernandez-Llamas, A., Vargas-Mendieta, M.,
recirculating raceway system for the production of marine shrimp. Aquac. Eng. 17,
Hernandez-Gurrola, J.A., Villarreal-Colmenares, H., 2018. Performance of a photo-
193–211.
heterotrophic, hypersaline system for intensive cultivation of white leg shrimp
Davis, D.A., Roy, L., Sookying, D., 2008. Improving the cost effectiveness of shrimp feeds.
(Litopenaeus vannamei) with minimal water replacement in lined ponds using a
271-280 pp. In: IX International Symposium on Aquaculture Nutrition. 24-27
stochastic approach. Aquac. Res. 49, 57–67.
November. Universidad Autónoma de Nuevo León, Monterrey, Nuevo León, México..
Moreno-Figueroa, L.D., Villarreal-Colmenares, H., Naranjo-Páramo, J., Vargas-
Ebeling, J.M., Timmons, M.B., Bisogni, J.J., 2006. Engineering analysis of the
Mendieta, M., Casillas-Hernández, L.M.R., Hernández-Llamas, A., 2019.
stoichiometry of photoautotrophic, autotrophic, and heterotrophic removal of
Bioeconomic modelling of the intensive production of whiteleg shrimp (Litopenaeus
ammonia-nitrogen in aquaculture systems. Aquaculture 257, 346–358.

6
D. Rodríguez-Olague et al. Aquaculture Reports 20 (2021) 100709

vannamei) in a photo-heterotrophic hypersaline system, with minimal seawater of the First Latin American Shrimp Culture Congress and Exhibition. Grupo Ferias,
replacement. Rev. Aquacult. 11 (3), 685–696. Eventos y Congresos. October 6–10, 1998. Panama City, Panama.
Moss, K.K., Moss, S.M., 2004. Effects of artificial substrate and stocking density on the Satanwat, P., Tran, T.P., Hirakata, Y., Watari, T., Hatamoto, M., Yamaguchi, T.,
nursery production of pacific white shrimp Litopenaeus vannamei. J. World Aquacult. Pungrasmia, W., Powtongsookf, S., 2020. Use of an internal fibrous biofilter for
Soc. 35 (4), 536–542. intermittent nitrification and denitrification treatments in a zero-discharge shrimp
Naegel, L.C.A., 2010. Management strategies to reduce operating costs in a commercial culture tank. Aquac. Eng. 88, 102041.
shrimp hatchery in NW Mexico. Aquac. Int. 18, 759–770. Silva, E., Silva, J., Ferreira, F., Soares, M., Soares, R., Peixoto, S., 2015. Influence of
Peacor, S.D., Shiesari, L., Werner, E.E., 2007. Mechanisms of nonlethal predator effect on stocking density on the zootechnical performance of Litopenaeus vannameiduring the
cohort size variation: ecological and evolutionary implications. Ecology 88, nursery phase in a biofloc system. Bol. Inst. Pesca, São Paulo. 41 (espe), 777–783.
1536–1547. Tierney, T.W., Fleckenstein, L.J., Ray, A.J., 2020. The effects of density and artificial
Persyn, H., Aungst, R., 2001. Global shrimp OP:200I-Preliminary report-nursery. Global substrate on intensive shrimp Litopenaeus vannamei nursery production. Aquac. Eng.
Aquacult. Advoc. 4 (4), 34–35. 89, 102063.
Peterson, J.J., Griffith, D., 1999. Intensive nursery systems. Global Aquacult. Advoc. 2, Valencia-Castañeda, G., Frías-Espericueta, M.G., Vanegas-Pérez, R.C., Chávez-
60–61. Sánchez, M.C., Páez-Osuna, F., 2019. Toxicity of ammonia, nitrite and nitrate to
Ponce-Palafox, J.T., Pavia, A.A., López, D.G.M., Arredondo-Figueroa, J.L., Lango- Litopenaeus vannamei juveniles in low-salinity water in single and ternary exposure
Reynoso, F., Castañeda-Chávez, M.D.R., Esparza-Leal, H., Ruiz-Luna, A., Páez- experiments and their environmental implications. Environ. Toxicol. Pharmacol. 70,
Ozuna, F., Castillo-Vargasmachuca, S.G., Peraza-Gómez, V., 2019. Response surface 103193.
analysis of temperature-salinity interaction effects on water quality, growth and Wasielesky Jr, W., Froes, C., Foes, G., Krummenauer, D., Lara, G., Poersch, L., 2013.
survival of shrimp Litopenaeus vannamei postlarvae raised in biofloc intensive Nursery of Litopenaeus vannamei reared in a biofloc system: the effect of stocking
nursery production. Aquaculture 503, 312–321. densities and compensatory growth. J. Shellfish Res. 32, 799–806.
Rajta, A., Bhatia, R., Setia, H., Pathania, P., 2020. Role of heterotrophic aerobic Williams, A.S., Davis, D.A., Arnold, C.R., 1996. Density-dependent growth and survival
denitrifying bacteria in nitrate removal from wastewater. J. Appl. Microbiol. 128, of Litopenaeus setiferus and Litopenaeus vannamei in a semi-closed recirculating
1261–1278. system. J. World Aquacult. Soc. 27 (1), 107–112.
Ray, A.J., Drury, T.H., Cecil, A., 2017. Comparing clear-water RAS and biofloc systems: Zar, J.H., 2010. Biostatistical Analysis. Prentice Hall, Upper Saddle River.
shrimp (Litopenaeus vannamei) production, water quality, and biofloc nutritional Zelaya, O., Davis, A., Rouse, D.B., 2007. The influence of Artemia and algal supplements
contributions estimated using stable isotopes. Aquac. Eng. 77, 9–14. during the nursery phase of rearing pacific white shrimp, Litopenaeus vannamei.
Samocha, T.M., Lawrence, A.L., 1998. Use of intensive nursery system in commercial J. World Aquacult. Soc. 38 (4), 486–496.
shrimp production: advantages and disadvantages. In: Jory, D.E. (Ed.), Proceedings