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Genetic manipulation in Bacillus subtilis using CRISPR/Cas9: a potential application in the control of viruses that affect shrimp farming View project
Evaluation of the effects of exposure to aflatoxin B1 through the ration on Litopenaeus vannamei subjected to low salinity View project
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Received: 18 April 2007 / Revised: 25 September 2007 / Accepted: 25 October 2007 / Published online: 12 December 2007
# Coastal and Estuarine Research Federation 2007
extension program. Rearing starts around the second week removing up to 30% of the nitrogen input through the
of December (austral spring time) using 1-month-old conversion of nitrate and ammonia into N2 (Fry 1987), but
postlarvae produced at the university facilities and stocked this is not the case for Patos Lagoon estuarine shallow
at a density of 20 shrimps m−2. Results show that the final waters, which show mean dissolved oxygen levels of 30 mg
yield of a standard pen of 0.3 ha and a density of 20 L−1 (Niencheski and Baumgarten 1997).
shrimps m−2 is 460 kg. This productivity provides the The intensity of the processes of consumption and
involved fishers with an additional monthly income of production depends mainly on factors related to the quality
approximately US$110 (Wasielesky et al. 2001). of the water, i.e., how rearing is carried out.
Water renewal rates estimated by Wasielesky (personal An empirical mixed box budget model for total nitrogen
communication) suggest that even under conditions of (TN) balance in shrimp rearing pens was designed and
lowest degrees of water flux the volume in the pen is elaborated using resources for modeling in the Stella
renewed with a daily rate of 320%. software, version 5.1, and the information used to run the
Shrimp is fed two times a day with PURINA feed. The model was obtained from the literature for the Pacific white
second time, in the late afternoon, is 60% of the daily ratio. shrimp (Litopenaeus vannamei). Model details are provided
Feed weight is a function of the shrimp biomass in the pen, in the Appendix.
which is 20% at the beginning and 2% at the end of the The release of nitrogen from the rearing pond was
rearing period. determined based on the rations of food supplied, water
To modelize the nitrogen consumption and production quality, and the growth rates of the reared specimens. The
processes occurring in a rearing pen, a conceptual diagram following assumptions were made; units are referred to
is proposed as shown in Fig. 3. weight in grams and total nitrogen:
The protein present in the shrimp’s feed is the largest
○ One kilogram of shrimp weight gain is equivalent to
source of nitrogen input into the system (Hargreaves 1998).
178.5 g protein and to 28.56 of N, or 2.85% of N on a
Part of the nitrogen entering the system is consumed by the
wet weight basis, according to L. vannamei body
shrimp and or is lost or not consumed. The unconsumed
composition (Boyd and Teichert-Coddington 1995).
feed usually represents 20% of that given (Wyban et al.
Following Holland et al. (1991), nitrogen in protein
1998), although for intensive rearing systems up to 50%
was estimated as 16% of crude protein equivalent to a
may be lost. Of the feed eaten, about 25% is converted into
proportion of 6.26 of crude protein to N.
growth and the rest is excreted through the gills and as
○ The protein content of the feed (Purina®) is 30%;
feces.
○ The feeding rate (FR) recommended by the feed
Phytoplankton and bacteria determine the assimilation
manufacturer (Purina®) for L. vannamei is:
capacity of nitrogen not assimilated by the shrimps.
Lorenzen et al. (1997) showed that phytoplankton may be FR ¼ 0:16 ðindividual weightÞ0:5
responsible for removing up to 87% of the dissolved
nitrogen present in a pond with low shrimp density and ○ Quantity of feed (QF) needed for a pen is calculated
54% with high density. This nitrogen uptake by phyto- multiplying FR by the biomass in the pen;
plankton limits the amount of nitrogen available for ○ There is a 15% loss of the feed given (NC), which is
sedimentation (Burford and Lorenzen 2004). Similarly, not consumed (Primavera 1993);
under anoxic conditions, bacteria are responsible for ○ The food ingested (IN) is:
IN ¼ QF NC
Entry Food offered ○ Of the total amount ingested, a fraction (%) is
Fertilization of the ponds
Natural water input digested (DI) and part is not. The digested food is
transformed into growth (GR) and excretion (EX), and
the undigested into feces (FC). Thus:
Growth of shrimp ,
Rearing
system Microorganism action
System s
assimilation
FC ¼ IN DI
Primary production capacity?
EX ¼ IN ðFC þ GRÞ
Harvest
Exit
Volatilization
Water renewal ○ 30% of the food ingested is transformed into growth
O.M. deposited (GR) (Montoya et al. 1999);
Fig. 3 Conceptual model of nitrogen consumption and production ○ The digestibility of the protein in the food is 90%
processes occurring in a rearing pen (O.M.: organic material) (Montoya et al. 1999);
218 Estuaries and Coasts: J CERF (2008) 31:215–222
○ At the end of the rearing process, there is a shrimp This adjustment (from 0.16 to 0.56) incorporated into the
mortality rate of 20%, considered as acceptable; model the natural food that is available in the environment
○ The initial weight of the shrimp (juveniles) is 0.35 g and that is present in the rearing process. After the new
(Wasielesky 2000); food ratio was adopted, the quantity of nitrogen that
○ The initial number of shrimp per pen (0.3 ha) is becomes available in the system and originates from the
60,000 (20 ind m−2). feces, gill excretion, and unconsumed food was recalculated
(Fig. 4).
To validate the model, water samples were collected in
Once the amount of total nitrogen released by a pen was
the vicinity of a single rearing pen located in the Ilha dos
known, a second stage of the simulation was carried out
Marinheiros (Fig. 1). The pen, with the standard area of
integrating the environment with the pen. To determine the
0.31 ha, was populated with 60,000 postlarvae of F.
appropriate number of pen enclosures for the Saco da
paulensis, which were fed on commercial feed. To
Mangueira, with minimal alterations to the water quality of
determine the total nitrogen in the water, samples were
the environment, a highly conservative scenario was
taken at four different distances from the pens (5, 15, 30,
adopted under the following assumptions:
and 100 m), the last and most distant one being the control
point during the 3-month period of shrimp rearing (one ○ Water circulation through the mesh (5 mm) of the
production cycle). Salinity, temperature, and the speed of pen is reduced by 15%;
the predominant currents were also measured. Analyses of ○ Speed of the water currents in the area is ≤0.01 cm
the total nitrogen were conducted at the hydrochemical s−1 (almost still water);
laboratory at FURG. ○ Maximum allowable concentration of total nitrogen
in the environment, resulting from rearing, is 0.07 mg
L−1. This figure is the average observed at the control
Results point for shrimp rearing in pens conducted at Ilha dos
Marinheiros (Patos Lagoon estuary).
Based on the above assumptions, a first version of the ○ All the nitrogen accumulated in the sediment during
model for total nitrogen balance in shrimp rearing pens was the rearing process is released to the environment at
devised and ran. However, it was considered to be harvesting time.
inadequate as the simulated growth rates for the shrimp
The model used to determine the number of pens in the
were lower than those observed in practice (Fig. 4). To
Saco da Mangueira can be seen in Fig. 5.
solve this problem, the ratio of food offered to the shrimp
Model validation was performed correlating predicted
was adjusted, so that:
(simulated) values of nitrogen input of the pen (1, 30, 60,
FR ¼ 0:56 ðindividualweightÞ0:5 and 90 days) with values observed in the field at 5 m
distance from the pen. A correlation coefficient of r=0.88
was found (Fig. 6). Predicted and observed nitrogen
concentrations were statistically and significantly correlat-
ed, indicating that model parameters and basic assumptions
FR = 0.56 x (individual weight) -0.5 were able to reproduce the processes of nitrogen produc-
12 600
tion, consumption, and dissipation into the environment.
Ind weight
Ind weight(g)
(g)adjusted
adjusted
Thus, new simulations were done to determine the number
10
Ind weight
Ind weight(g)
(g) of pens the environment could hold.
To estimate the allowable number of pens in the Saco da
8 400
Mangueira, a cautious approach was followed. Simulations
Biomass (kg)
Weight (g)
6
were carried out with a water current speed of 0.01 cm s−1,
which is lower than the actual value observed in the
4 200
environment. For instance, current measurements taken
during February at the Saco do Justino (Fig. 1)—a bay with
2 lower hydrodynamics than the Saco da Mangueira—
showed values between 0 and 3 cm s−1.
0 0 Considering a current of 0.01 cm s−1 and a shrimp
density of 20 ind m−2, the simulations showed that 86 pen
0 9 18 27 36 45 54 63 72 81 90
Rearing days
Fig. 4 Estimate of growth of shrimp in pen, using a mortality rate of
enclosures could be installed in the area before the
20%, the feeding rate (FR) recommended by Purina® (squares) and established limit of 0.07 mg L−1 of N-NT was reached.
the adjusted ratio used in the model (circles) However, when the water current speeds were reduced even
Estuaries and Coasts: J CERF (2008) 31:215–222 219
Fig. 8 Effect of the number of pens on N-NT emission for the Saco
da Mangueira. A and B represent a scenario of 70 and 200 pen
Fig. 6 Results of the correlation analysis of the data produced by the enclosures, respectively, with 20 shrimps m−2 and a water current
model and that observed speed of 0.01 cm s−1
220 Estuaries and Coasts: J CERF (2008) 31:215–222
Fig. 9 Simulation with 70 pens (20 shrimp m−2) and with no water
circulation
Discussion
Appendix References
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aquaculture in developing countries. ICLARM Conference penaeus paulensis (Decapoda, Penaeidae) no estuário da Patos
Proceedings. Lagoon: Efeitos de parâmetros ambientais e manejo de cultivo.
Piedrahita, R. 2003. Reducing the potential environmental impact of Tese de doutorado, Fundação Universidade Federal do Rio
tank aquaculture through intensification and recirculation. Aqua- Grande. Rio Grande, Brasil.
culture 226:35–44. Wasielesky, W., L.H. Poersch, L. Jensen, and A. Bianchini. 2001.
Primavera, J.H. 1993. A critical review of shrimp pond culture in the Effect of stocking density on pen reared pink shrimp Farfante-
Philippines. Reviews in Fisheries Science 1(2):151–201. penaeus paulensis (Perez-Farfante, 1967) (Decapoda, Penaeidae).
Rosenberry, B. 1998. World shrimp farming. Shrimp News International. Nauplius 9(2):163–167.
Velasco, M., A.L. Lawrence, and W.H. Neill. 1998. Development of a Wyban, J.A., J.N. Sweeney, and R. Kanna. 1998. Shrimp yields and
static-water ecoassay with microcosm tanks for postlarval economic potential of intensive round pond systems. Journal of
Penaeus vannamei. Aquaculture 161:9–87. World Aquaculture Society 19:210–217.