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Effect of different concentrations of potassium and magnesium on

performance of Litopenaeus vannamei postlarvae reared in low-salinity water
and a biofloc system

Article  in  Journal of Applied Aquaculture · October 2018

DOI: 10.1080/10454438.2018.1536009

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Effect of different concentrations of potassium

and magnesium on performance of Litopenaeus
vannamei postlarvae reared in low-salinity water
and a biofloc system

Simão Zacarias, Rodrigo Schveitzer, Rafael Arantes, Helena Galasso, Isabela

Pinheiro, Carlos Espirito Santo & Luis Vinatea

To cite this article: Simão Zacarias, Rodrigo Schveitzer, Rafael Arantes, Helena Galasso,
Isabela Pinheiro, Carlos Espirito Santo & Luis Vinatea (2018): Effect of different concentrations of
potassium and magnesium on performance of Litopenaeus�vannamei postlarvae reared in low-
salinity water and a biofloc system, Journal of Applied Aquaculture

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JOURNAL OF APPLIED AQUACULTURE
https://doi.org/10.1080/10454438.2018.1536009

Effect of different concentrations of potassium and

magnesium on performance of Litopenaeus vannamei
postlarvae reared in low-salinity water and a biofloc
system
Simão Zacariasa, Rodrigo Schveitzerb, Rafael Arantesc, Helena Galassoc,
Isabela Pinheiroc, Carlos Espirito Santoc, and Luis Vinateac
a
Institute of Aquaculture, University of Stirling, Stirling, Scotland, UK; bDepartment of Marine Sciences,
Federal University of São Paulo (UNIFESP), Santos, São Paulo, Brazil; cCentro de Ciências Agrárias,
Departamento de Aquicultura, Laboratório de Camarões Marinhos, Universidade Federal de Santa
Catarina, Florianópolis, Brazil

ABSTRACT KEYWORDS
Effect of K+ and Mg2+ in water on the performance of Litopenaeus Inland water; ionic
vannamei using bioflocs was evaluated. Control had water with K: concentration; ions; shrimp
Mg (1:4.57) and other treatments (K:Mg of 1:5.4, 1:4.65, and 1:3.77),
both with salinity of 4 g/L. Initial mean weight of shrimp was
0.08 ± 0.007 g. No difference in shrimp growth performance and
survival was observed (P > 0.05) except in the control. Our results
suggest that it is possible to culture postlarvae of Litopenaeus
vannamei in low-salinity water with bioflocs when the water has
an initial potassium concentration of 30.90 ± 8.5 mg/L and mag-
nesium of 167.0 ± 3.9 mg/L.

Introduction
Pacific white shrimp (Litopenaeus vannamei) inhabits natural environments
with salinity ranging from 0 to 60 g/L; thus, it has high potential for culture in
marine or inland water (Mcgraw 2002; Davis, Samocha, and Boyd 2004) . In
China, Thailand, Vietnam, Ecuador, Mexico, the United States, and Brazil,
culture of this species in low-salinity inland waters is a common practice
(Boyd and Thunjai 2003; Boyd, Thunjai, and Boonyaratpalin 2002; Roy et al.
2010). Water used to culture Pacific white shrimp in low salinity comes from
different sources and generally has ionic deficiencies, mainly of ions considered
necessary for L. vannamei culture (Boyd 2006; Roy, Davis, and Nguyen 2009;
Saoud, Davis, and Rouse 2003; Zhu et al. 2006). Therefore, it might be necessary
to correct ionic water profile before and during culture (Roy, Davis, and Nguyen
2009; Roy et al. 2007b)
Bicarbonates and carbonates are considered fundamental in low-salinity
marine shrimp culture; however, sulfate, magnesium, calcium, potassium,

CONTACT Simão Zacarias simaozacarias@yahoo.co.uk; simao.zacarias1@stir.ac.uk Institute of Aquaculture,

University of Stirling, Stirling FK9 4LA, Scotland, UK
© 2018 Taylor & Francis
2 S. ZACARIAS ET AL.

sodium, and chloride are the most important (Boyd 2006; Boyd and Thunjai
2003). According to Boyd and Thunjai (2003) and Davis, Samocha, and Boyd
(2004), minimum concentrations of these ions necessary for L. vannamei
culture in low-salinity water are not known, but it is considered that marine
shrimp survive and grow well when ionic proportion of low-salinity water is
similar to diluted sea water of the same salinity.
According to McGraw (2002) and Boyd (2006), chloride, sodium, sulfate,
calcium, magnesium, and potassium play a role in L. vannamei osmoregulation.
Potassium and magnesium are essential for culture success in low-salinity waters as
they are involved in growth, survival, and osmrregulation (McGraw and Scarpa
2003; Roy et al. 2007b). One of the problems for the success of L. vannamei culture
in low-salinity waters is lack of potassium and magnesium (Roy, Davis, and
Nguyen 2009). Potassium is essential for activation of Na+-K+-ATPase enzyme,
an important component of extracellular volume regulation (Roy et al. 2007a).
Magnesium acts as a cofactor of Na+-K+-ATPase enzyme and of proteins, lipids,
and carbohydrates (Roy et al. 2007a). During the molting process shrimp absorb
high quantities of magnesium and calcium for mineralization of their exoskeleton
(Boyd and Tucker 1998).
Biofloc is a heterogenic mixture of aggregates, composed of bacteria, phyto-
plankton, protozoan, filamentous organisms, algae, organic particles, colloids,
organic polymer, dead cells, and others components (Avnimelech 2009; De
Schryver et al. 2008). Biofloc formation occurs by adding a supplemental organic
carbon source (other than feed), the main function of which is to increase the
carbon/nitrogen ration of organic substrate and stimulate heterotrophic bacterial
growth, responsible for ammonia absorption (Avnimelech 2009; Emerenciano,
Gabriela, and Gerard 2013; Hargreaves 2013). Few studies have applied biofloc
technology to marine shrimp culture in inland low-salinity waters. Advantages of
using this technology in a low-salinity environment would be similar to those
reported in marine environment culture: improving water quality, increasing
natural food availability, improving feed conversion, and pathogen biocontrol
(Avnimelech 2009). Additionally, it would contribute to reduce use of water and
salinization of soils, aquifers, and rivers, and avoid eutrophication of surrounding
water bodies (Boyd 2006).
Lack of information on the effect of ions on shrimp production in a
biofloc system may compromise the application of this technology in inland
low-salinity waters. According to De Schryver et al. (2008), biofloc has also
some ions, but the composition varies highly. Addition of ions to water with
bioflocs contributes to their stabilization (Avnimelech 2009). Lodão (2009)
reports that growth and development of microorganisms associated with
organic substrates, such as heterotrophic bacteria, which compose bioflocs,
are affected by inorganic nutrient availability. Furthermore, Luo et al. (2013)
and Eldyasti, Nakhla, and Zhu (2013) say that divalent cations, such as Ca2+
and Mg2+, help biofloc formation. Considering that shrimp as well as bacteria
 JOURNAL OF APPLIED AQUACULTURE 3

can use potassium and magnesium, and in a biofloc system shrimp stocking
density and quantity of bacteria are high, it is possible that the concentration
of ions used in semi-intensive culture systems (Roy et al. 2007b) is not
enough to maintain shrimp performance in a biofloc system. In this case, it
might be necessary to increase the concentrations of these ions. Our work
evaluated the effect of different concentrations of potassium and magnesium
in water on performance of Litopenaeus vannamei postlarve reared in low
salinity and a biofloc system.

Materials and methods

This study was conducted in the Marine Shrimp Laboratory (LCM) of the
Federal University at Santa Catarina. Litopenaeus vannamei postlarvae used in
the experiment had mean weight of 0.08 ± 0.007 g and were produced by
LCM. Prior to the experiment, shrimp were acclimated to experimental low-
salinity water (4 g/L). In this process, postlarvae (PL24-28) were stocked into
two fiberglass tanks each with 800 L of usable volume, and original salinity
(32 g/L) was maintained for the first 24 hours. After that, 50% of the tank
volume was replaced with dechlorinated freshwater for three days until reach-
ing salinity of 4 g/L. During acclimation, postlarvae were fed a specific feed
(EPAC PL®—50% protein and 10% lipid; EPAC XL®—50% protein and 10%
lipid) nine times a day, and water temperature was maintained in 26.0ºC,
dissolved oxygen above 7.0 mg/L, and pH close to 8.0. Shrimp remained 6
more days postacclimation and before transferring them to experimental units.
The experiment was entirely randomly designed with four treatments and
three replications (12 experimental units), in which L. vannamei PLs were
cultured during 32 days at a density of 220/m2. Shrimp were cultured in
artificial brackish water (salinity of 4.0 g/L) with different concentrations of
potassium and magnesium, and the target ratios (mg/L) were 35:150 (lower
than the control) for the first treatment, 45:170 (similar to the control) for
the second, and 55:190 (higher than the control) for the last one. Potassium
and magnesium concentrations were selected considering that their require-
ment in a biofloc system would be most likely higher or less likely lower than
normal (control). Artificial brackish water was prepared by adding potassium
chloride (KCl—potassium source, 99% pure), heptahydrated magnesium
chloride—(MgCl2.7H2O—magnesium source, 99% pure), and calcium carbo-
nate (CaCO3—calcium source, 98% pure) to dechlorinated fresh water. After
that the solution was homogenized for 12 hours to ensure that all chemicals
were completely dissolved, sodium chloride (99.5% pure) was added to
correct salinity to 4.0 g/L. This salinity was adopted in this study because
Maicá, de Borba, and Wasielesky (2012) obtained good productivity of L.
vannamei cultured in a biofloc system and marine reconstituted water with
salinity of 4.0 g/L. As a control, another group of PLs was cultured in marine
4 S. ZACARIAS ET AL.

reconstituted water with salinity of 4.0 g/L, which was prepared by mixing
dechlorinated fresh water and marine water with salinity of 32.0 g/L. Analysis
of potassium, magnesium, calcium, sodium, and chloride was done one day
before stocking the experimental units (Table 2), and the average concentra-
tions of potassium and magnesium were close to desired concentrations.
Water samples used for these analyses were not filtered and were performed
through Alfakit® Ionic Balance Polikit, which follows the methodology of
American Public Health Association (APHA, 2005). After the beginning of
the experiment, these ions were analyzed twice a week (Alfakit®); however,
water samples had to be filtered from this point due to presence of flocs.
Experimental units consisted of circular fiberglass tanks of 1,000 L (1 m2
surface area), which were kept in a greenhouse and received natural lighting.
A central aeration ring (aero-tube™) was used to aerate the system and
maintain bioflocs in suspension, and submersible heaters controlled the
temperature. Shrimp were fed four times a day with feed of 40.0% crude
protein and 7.5% lipids (Guabi® “Potimar 40PL”). The quantity of feed
offered was equal in all treatments, and the feeding rate was 3.5% of shrimp
biomass per day, but it was adjusted according to weekly animal growth (Van
Wyk and Scarpa 1999). Cane molasses addition was calculated according to
Avnimelech (1999) assuming that 20 g of carbohydrate are necessary to
convert 1 g of ammonia in bacterial biomass. Due to increased concentration
of ammonia during the experiment, cane molasses was added until the week
before the last one. Water was not renewed during the 32 days of study;
however 25 L of dechlorinated fresh water was added in each tank every week
to compensate for evaporation without affecting the experimental salinity.
During the experiment, temperature and dissolved oxygen were measured
twice a day using a digital oxymeter (YSI Pro20). Salinity was determined twice
a week with a salinity meter (YSI 3200), and pH was analyzed daily with a
portable pH-meter (YSI 100). Water samples were collected twice a week from
each tank to determine alkalinity (APHA, 2005), total suspended solid (APHA,
2005), total ammonia (Grasshoff, Ehrhardt, and Kremling 1983), nitrite
(Baumgarten, Rocha, and Niencheski 1996), nitrate (HACH method 8039,
cadmium reduction), and phosphate (Baumgarten, Rocha, and Niencheski
1996). Survival—100 x (final shrimp number / initial shrimp number), final
mean weight—(final biomass / final shrimp number), total weight gain (final
mean weight – initial mean weight), final biomass (final shrimp number x final
mean weight), and specific growth rate—100 x (ln final weight – ln initial
weight) / time in days—were used to assess the culture performance. Shrimp
were sampled once a week and after weighing were returned to their respective
tanks.
One-way ANOVA followed by Tukey’s test (Zar 2010) was used to compare the
treatments with a significance level of 0.05. Normality and homoscedasticity were
tested using Shapiro-Wilk and Levene tests respectively. A Kruska-Walls test was
 JOURNAL OF APPLIED AQUACULTURE 5

used in the absence of these characteristics. Coefficient of determination was used

to understand the relationship between the amount of molasses added in each
treatment and potassium, magnesium, and calcium concentrations during the
experiment.

Results
There were no significant differences among treatments for water quality
variables (P > 0.05) (Table 1). Ion concentrations analyzed one day before,
during, and at the end of the experiment, and coefficient of determination
between cane molasses and potassium, magnesium, and calcium concentra-
tions are shown in Table 2. Values of ions concentrations measured one day
before the experiment represent the actual values for the study. Mean
potassium and calcium at onset of the experiment are lower than the final
values, while for other ions they are higher. This reveals that potassium and
calcium concentrations increased over time, while other ions dropped in all
treatments. Growth performance parameters and survival are presented in
Table 3. Survival of the control was significantly lower than treatment with
potassium and magnesium level of 35:150 and 55:190 (P < 0.05) but equal to
45:170 (P > 0.05). Total weight gain, mean final weight, specific growth rate
(SGR), and final biomass were similar in all treatments except the control.

Discussion
Dissolved oxygen, temperature, and pH were within recommended ranges for
Litopenaeus vannamei culture (Van Wyk and Scarpa 1999; Whetstone et al. 2002).
Mean un-ionized ammonia concentrations of all treatments were below LC50

Table 1. Water quality variables in L. vannamei culture in low salinity and a biofloc system with
different concentrations of potassium and magnesium in water. Values are mean ± standard
error. n = 3.
Treatments
Variables K:Mg (1:5.4) K:Mg (1:4.65) K:Mg (1:3.77) Control (1:4.57)
Oxygen morning (mg/L) 7.4 ± 0.1 7.4 ± 0.0 7.4 ± 0.1 7.4 ± 0.1
Oxygen afternoon (mg/L) 7.0 ± 0.0 7.1 ± 0.1 7.1 ± 0.1 7.0 ± 0.1
Temperature morning (°C) 28.4 ± 0.2 28.3 ± 0.4 28.4 ± 0.2 28.1 ± 0.2
Temperature afternoon (°C) 30.2 ± 0.2 29.9. ± 0.5 29.9 ± 0.2 29.8 ± 0.2
pH 8.4 ± 0.0 8.4 ± 0.0 8.4 ± 0.0 8.1 ± 0.1
Salinity (g/L) 4.2 ± 0.0 4.3 ± 0.0 4.2 ± 0.0 4.2 ± 0.1
Alkalinity (mg/L CaCO3) 154.0 ± 10.1 161.2 ± 10.3 162.7 ± 2.9 137.3 ± 3.1
TA-N (mg/L) 1.4 ± 0.1 1.4 ± 0.1 1.5 ± 0.2 1.3 ± 0.5
NH3-N (mg/L) 0.3 ± 0.0 0.2 ± 0.0 0.3 ± 0.0 0.1 ± 0.0
NO2-N (mg/L) 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0
NO3-N (mg/L) 5.2 ± 0.6 5.0 ± 0.2 5.0 ± 0.1 4.9 ± 0.8
PO43-P (mg/L) 0.6 ± 0.0 0.6 ± 0.0 0.5 ± 0.0 0.7 ± 0.0
Total suspended solids (mg/L) 246.1 ± 12.7 312.2 ± 15.0 330.0 ± 15.6 259.1 ± 8.2
Note. Variables were analyzed by ANOVA. There was no significant difference among treatments (P > 0.05).
 6

Table 2. Concentration and ion ratios above the line are initial values (one day before the experiment) (n = 3). Below the line are mean values and ions ratios
during the experiment and coefficient of determination (n = 3). Means in brackets are final values (last day of the experiment) (n = 3). Values are mean ± standard
error.
Treatments
Ions K:Mg (1:5.4) K:Mg (1:4.65) K:Mg (1:3.77) Control (1:4.57)
K+ (mg/L) 30.9 ± 4.9 44.7 ± 6.3 56.7 ± 5.3 45.0 ± 8.6
S. ZACARIAS ET AL.

(81.7 ± 11.9) (95.1 ± 31.9) (99.4 ± 10.4) (106.7 ± 23.9)

Mg2+ (mg/L) 167.0 ± 2.3 208.1 ± 6.7 213.9 ± 8.1 205.7 ± 15.4
(134.3 ± 10.1) (142.2 ± 39.8) (163.3 ± 12.2) (163.8 ± 9.9)
Ca2+ (mg/L) 35.2 ± 2.6 39.2 ± 5.3 37.9 ± 5.1 43.2 ± 1.7
(69.9 ± 21.0) (72.3 ± 22.1) (88.8 ± 11.6) (117.33 ± 11.7)
Cl– (mg/L) 3005.7 ± 23.7 2989.0 ± 71.0 3029.3 ± 23.7 2958.3 ± 62.6
(2082.7 ± 333.8) (1893.3 ± 520.7) (2224.7 ± 155.2) (2224.7 ± 23.7)
Na+ (mg/L) 1660.9 ± 13.1 1647.9 ± 39.2 1674.0 ± 13.07 1634.8 ± 34.6
(1150.9 ± 184.5) (1046.3 ± 287.7) (1229.3 ± 85.7) (1229.3 ± 13.0)
Na:K 53.7:1 36.9:1 29.5:1 36.3:1
Ca:K 1.1:1 0.9:1 0.7:1 1:1
Mg:Ca 4.7:1 5.3:1 5.6:1 4.8:1
K+ (mg/L) 57.5 ± 2.0 75.0 ± 5.6 84.4 ± 4.0 75.0 ± 4.8
Mg2+ (mg/L) 145.5 ± 1.9 170.3 ± 7.9 182.8 ± 2.4 170.5 ± 4.1
Ca2+ (mg/L) 58.9 ± 2.3 64.5 ± 1.1 66.1 ± 1.0 81.3 ± 4.2
Cl– (mg/L) 2627.0 ± 64.4 2584.4 ± 39.1 2634.1 ± 18.8 2541.8 ± 33.5
Na+ (mg/L) 1451.7 ± 35.6 1428.1 ± 21.6 1455.6 ± 10.4 1404.6 ± 18.5
Na:K 25.2:1 19.0:1 17.2:1.0 18.7:1.0
Ca:K 1.0:1.0 0.9:1.0 0.8:1.0 1.1:1.0
Mg:Ca 2.5:1.0 2.6:1.0 2.8:1.0 2.1:1.0
R2 K+ (%) 83.53 72.60 65.23 68.25
R2 Mg2+ (%) 50.97 32.06 53.62 45.48
R2 Ca2+ (%) 94.83 93.24 91.23 91.99
Note. R2 — Coefficient of determination between cane molasses and ion.
 JOURNAL OF APPLIED AQUACULTURE 7

Table 3. Performance of L. vannamei cultured in low-salinity water with bioflocs and different
concentrations of potassium and magnesium. Mean ± Standard Error. n = 3.
Treatments
Parameters K:Mg (1:5.4) K:Mg (1:4.65) K:Mg (1:3.77) Control (1:4.57)
Survival (%) 87.5 ± 3.3b 81.2 ± 3.6ab 89.8 ± 0.4b 65.0 ± 6.17a
Total weight gain (g) 0.7 ± 0.1b 0.6 ± 0.1b 0.6 ± 0.0b 1.5 ± 0.2a
Mean final weight (g) 0.8 ± 0.1b 0.7 ± 0.0b 0.7 ± 0.0b 1.7 ± 0.2a
SGR (%) ** 7.0 ± 0.2b 6.4 ± 0.5b 6.6 ± 0.1b 9 ± 0.4a
Final biomass (Kg/m3) 0.15 ± 0.0b 0.12 ± 0.0b 0.14 ± 0.0b 0.22 ± 0.0a
Note. Variables were analyzed by ANOVA. Letter that are different in same row are significantly different
(P < 0.05).
*Initial mean weight of shrimp (g): 0.08 ± 0.007.
**Specific growth rate.

reported by Lin and Chen (2001) in low-salinity water. Likewise nitrite, nitrate, and
alkalinity were suitable for L. vannamei culture (Van Wyk and Scarpa 1999). Total
suspended solids did not exceed 500 mg/L, which is the level considered optimum
for culture of L. vannamei with bioflocs (Schveitzer et al. 2013).
The actual values of potassium and magnesium at the onset of the experiment
were close to targeted concentrations. Therefore they were not adjusted. Roy et al.
(2007a) used similar methodology. Potassium and calcium concentrations rose in
all treatments during the experimental period. This was not expected and was most
probably due to the cane molasses input, as the coefficients of determination
showed a stronger relationship between amount of added molasses and rise of
potassium and calcium in water. Magnesium concentration decreased in all treat-
ments, and the coefficient of determination confirmed that magnesium concen-
tration was not directly related to cane molasses addition. The reduction of
magnesium may be due to the low concentrations in molasses (Curtin 1973) in
relation to the need of this ion by the shrimp and microbial community.
Shrimp use potassium and magnesium in water for osmoregulation, growth,
and survival (McGraw and Scarpa 2003; Roy et al. 2007b). Microorganisms also
use those ions in water (Luo et al. 2013), including heterotrophic bacteria that
compose bioflocs (Lodão 2009). Addition of calcium and magnesium in water
contribute to the formation of bioflocs (Avnimelech 2009; Eldyasti, Nakhla, and
Zhu 2013; Luo et al. 2013). Consumption of potassium, magnesium, and calcium
by shrimp and microorganisms would lead to depletion of the ions in a closed
system. Therefore Boyd, Thunjai, and Boonyaratpalin (2002) and Roy et al. (2010)
suggested that shrimp producers should monitor concentrations of ions in water
before and during culture in order to adjust them to normal levels using muriate of
potassium (KCl) or potassium sulphate (K2SO4) as a potassium source, dolomitic
limestone (MgCO3xCaCO3) or magnesium suphate (Mg2SO4) as a magnesium
source, and agriculture limestone (CaCO3 or MgCO3xCaCO3) as a calcium source.
Our results indicate that this adjustment might not be required for some ions such
as potassium or calcium when using a biofloc system fertilized with cane molasses.
It seems that potassium and calcium consumption by shrimp and microorganisms
8 S. ZACARIAS ET AL.

of biofloc is not enough to deplete their concentration in water. Our initial

management strategy was to replace consumed ions during culture, but the
increase in the level of potassium and calcium impeded us to add ions. Our
research is the first one to report an increase in potassium and calcium concentra-
tions during shrimp culture with a biofloc system. Farmers using this technology
with cane molasses in inland areas are required to get the ionic composition of their
organic carbon source to better manage ionic balance in a biofloc system. In
addition, future studies should do ionic composition of cane molasses to full
understand the dynamic of ions in a biofloc system.
In a study with low-salinity clear water (4 g/L), Roy et al. (2007a) observed
that an increase in potassium concentration (5 to 40 mg/L) improved survival
and growth of L. vannamei. Survival of Pacific white shrimp postlarvae was
positively correlated with magnesium and potassium levels in clear low-
salinity water postacclimation (Saoud, Davis, and Rouse 2003). Davis et al.
(2005) also found a positive correlation between PL survival following accli-
mation to low-salinity water and level of potassium. Purushothaman et al.
(2014) observed total mortality of P. monodon PLs in tanks without supple-
mentation of potassium in brackish water but not in supplemented groups
with different levels. Growth performance and survival results found in the
present study do not corroborate previous studies, as no advantages of
increasing levels of potassium and magnesium in water with biofloc was
observed.
Boyd and Thunjai (2003), Davis, Samocha, and Boyd (2004) and Roy,
Davis, and Saoud (2006) state that to guarantee good survival and growth of
shrimp, ratios of ions in water should be closer to those found in marine
water. Lack of an adequate Na:K ratio resulted in low activity and death of
juvenile shrimp of L. vannamei at a salinity of 30 g/L (Zhu et al. 2004). The
same authors also reported that shrimp growth improved when Na:K is
nearly 40–43:1. Esparza-Leal et al. (2009) observed a positive impact of Na:
K (29.5:1) and Ca:K (16.4:1) ratios on survival and growth of L. vannamei.
Roy et al. (2007a) found improvement from 5 to 40 mg/L of potassium
because their Na:K ratio was closer to marine water in 40 mg/L concentration
(29:1). They also observed that concentration of magnesium above 20 mg/L
was enough to facilitate good survival and weight gain of L. vannamei, as
above this concentration Mg:Ca2 ratios were similar to normal seawater
(3.1:1). Pan, Luan, and Jin (2006) found significant survival rates and weight
gain of Marsupenaeus japonicus postlarvae when Na:K ratios were between
20–30:1 and Mg:Ca2 ratio of 4.5. Growth and survival of L. vannamei PLs
reared at the Na:K ratio of 40:1 were higher than at the ratio of 120:1 at 20ºC,
and the survival rate only at 24ºC (Perez-Velazquez et al. 2012). In the
current study the effect of experimental treatments was affected as potassium
and calcium increased over the experimental period and ions ratios of Na:K,
Ca:K and Mg:Ca have changed over time. This change might have made ion
 JOURNAL OF APPLIED AQUACULTURE 9

ratios of all treatments to be similar to normal seawater. Therefore, all

experimental treatments resulted in the same effect on shrimp performance.
Survival in the control was relatively lower (65.0 ± 5.7%), and a specific
reason was not found for that. The control had marine reconstituted
water in which the ions ratio was similar to normal seawater (Table 2),
as also demonstrated by Roy et al. (2007a), and this would theoretically
give similar or better survival. Based on our data, there is no clear
explanation for the lower survival rate in the control. However, survival
in the control was similar to the treatment with potassium and magne-
sium ratio 1:4.65. This similarity was observed because both groups had
an identical initial ionic ratio of Na+:K+, Mg2+:Ca2+ and K+:Mg2+ (Table
2), which suggests that ionic ratios are also important in a biofloc system
where heterotrophic microorganisms consume ions as well. Furthermore,
Maicá, de Borba, and Wasielesky (2012) observed similar survival to our
control (72 ± 16.6%) using marine reconstituted water with the same
salinity (4 g/L) and biofloc system, which indicates that there might be
other factors, such as other ions in the marine reconstituted water that
may affect the survival of the shrimp.
Other growth performance parameters in the control were significantly
affected by lower survival. Final weight (1.7 ± 0.4 g) and total weight gain
(1.5 ± 0.0 g) were significantly higher than other treatments, but they
were similar to those obtained by Maicá, de Borba, and Wasielesky (2012),
1.04 ± 0.07 g and 1.70 ± 0.07 g respectively. However, specific growth
rates were higher than those found by these authors (5.23 ± 0.09%). In
treatments with addition of ions, concentrations and ion ratios of potas-
sium and magnesium were different from the beginning to the end of the
experiment; however, survival, final weight, total weight gain, specific
growth rate, and final biomass were similar throughout. This might be
associated with the level of potassium and magnesium concentrations in
water, as the mean values during the experiment were above the mini-
mum required to culture Pacific white shrimp in low-salinity water (Boyd
and Thunjai 2003; Davis, Samocha, and Boyd 2004). Additionally, increas-
ing potassium and calcium has changed the ratio of Na+:K+, Mg2+:Ca2+
and Mg2+:K+ over time, and this probably allowed all treatments to have
similar ion ratios to seawater; therefore the effect was equal.

Conclusion
Our results demonstrate that it is possible to culture PLs of Litopenaeus
vannamei in a nursery using low-salinity water (4 g/L) with a biofloc
system and when the water has an initial mean potassium concentration
of 30.9 ± 8.5 mg/L and magnesium of 167.0 ± 3.9 mg/L. However, there is
a need to verify other factors such as ionic water balance, since the
10 S. ZACARIAS ET AL.

control exhibited lower survival. In addition, studies evaluating the budget

of important ions like potassium, calcium, and magnesium are encouraged
to further understand their dynamics in a biofloc system and should
include a clear-water system as the control.

Acknowledgments
Thanks to Robyn Livesey-Shilland for proofreading the English language.

Disclosure statement
No potential conflict of interest was reported by the authors.

Funding
This work was supported by the Conselho Nacional de Desenvolvimento Científico e
Tecnológico [151032/2012-2]; Southern Ocean and Education Development (SOED)—
Canada program for the master’s scholarship and funds for research (Zacarias) .

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