Ethical statement
The study was done by employing the protocol of CPCSEA (Committee for the Purpose of Control and Supervision of Experiments on Animals), Ministry of Environment and Forests (Animal Welfare Division), Govt. of India on welfare and utilization of animals in scientific research. This study was approved by the ethical committee of ICAR-Central Institute of Fisheries Education, Mumbai, India.
Experimental animals and design
The experiment was carried out from June to November, 2023. Juveniles of
C.
striata with a mean weight of 4.74± 0.09 g were acquired from an aqua consultant in Andhra Pradesh, India. After being transported to the ICAR-Central Institute of Fisheries Education (CIFE) regional centre, Rohtak, Haryana, India, the fish were acclimated in tanks containing freshwater for 15 days. After acclimatization, the juveniles with a mean weight of 6.21±0.03 g were progressively changed to salinities of 12 and 14 ppt, respectively, by adding 1 ppt every day within 15-day intervals. Throughout the acclimatization period, the fish were provided with a Growel commercial feed with 42% crude protein (CP), 3% crude fibre (CF), 8% crude lipid (CL) and 12% moisture content twice daily @ 5% biomass. The tanks were provided with proper aeration to maintain an appropriate DO level.
After 15 days, twenty-four
C.
striata juveniles with mean biomass of (8.31±0.05 g) were randomly released in eight different experimental units for the Se experiment
viz., (12 ppt) 0 mgSe (0 mg sodium selenite (inorganic form of Se) (Na
2SeO
3)/kg of diet), (12 ppt) 0.1 mgSe (0.1 mg Na
2SeO
3/kg of diet), (12 ppt) 0.3 mgSe (0.3 mg Na
2SeO
3/kg of diet), (12 ppt) 0.5 mgSe (0.5 mg Na
2SeO
3/kg of diet) were reared in 12 ppt salinity whereas, (14 ppt) 0 mgSe (0 mg Na
2SeO
3/kg of diet), (14 ppt) 0.1 mgSe (0.1 mg Na
2SeO
3/kg of diet), (14 ppt) 0.3 mgSe (0.3 mg Na
2SeO
3/kg of diet), (14 ppt) 0.5 mgSe (0.5 mg Na
2SeO
3/kg of diet) were reared in 14 ppt salinity in triplicates following a 2 ´ 3 factorial design for 150 days. Similarly, the fish were randomly stocked in another eight experimental units for the Tau experiment
viz., (12 ppt) 0 gTau (0 g Tau/kg of diet), (12 ppt) 5 gTau (5 g Tau/kg of diet), (12 ppt) 10 gTau (10 g Tau/kg of diet), (12 ppt) 15 gTau (15 g Tau/kg of diet) were reared in 12 ppt salinity whereas, (14 ppt) 0 gTau (0 g Tau/kg of diet), (14 ppt) 5 gTau (5 g Tau/kg of diet), (14 ppt) 10 gTau (10 g Tau/kg of diet), (14 ppt) 15 gTau (15 g Tau/kg of diet) were reared in 14 ppt salinity in triplicates employing a 2 x 3 factorial design for an experimental period of 150 days. Circular fibre-reinforced plastic (FRP) tanks with a capacity of 400 L (1.0 m x 0.5 m) were used for experiments and 250 L water volume was maintained in every tank with proper aeration during the experiment trial. The
C.
striata were hand-fed two times daily (08:30 and 16:30 hrs.) @ 5% biomass to their satiation level. 30% water was exchanged regularly to eradicate the faecal matter and uneaten feed.
Preparation of experimental diet
The feed employed in this study is a commercial diet from Growel, India, having a size of 1.2 mm with 42.0% CP, 8.0% CL, 3.0% CF and 12.0% moisture and with this, Se and Tau were separately added through the surface coating method using the Unispray equipment (S. B. Panchal and Company, Mumbai). The feed additive Se was purchased from Merck Schuchardt OHG, Germany and Tau was procured from Virion Enterprises, Mumbai. As per the experimental requirement, Se and Tau were weighed and diluted in the commercial coating gel (Turbogel, Guybro Chemical) (50-70 ml/kg feed) according to manufacturer instructions and stirred for homogeneous distribution. After the even coating, the feed was dried at room temperature for at least one day before feeding. The control diet was coated with the same coating gel to avoid an experimental error.
Preparation of fillets
After being cultured for 150 days,
C.
striata were euthanized by cervical dislocation following hypo-thermal stunning and placed in a cooler containing ice-cold water at 0-4
oC. After being washed using freshwater, the grown
C.
striata were segregated into ten samples for every replicate and brought to the processing lab on crushed ice. To investigate fillet quality, a competent fish processor filleted the fish.
Colour analysis
Using a CR-300 chromometer (Minolta, Osaka, Japan), the colour of the
C.
striata fillet was assessed in both raw and cooked forms. The apparatus measures the CIE-L* brightness of the
C.
striata raw and cooked fillet (Lx = 0 for black, L* = 100 for white), a* redness (a* > 0) and b* yellowness (b* > 0), with the chroma ((a*)2 + (b*)2)1/2 being the position in the colour space. The colour was recorded above the lateral line at three points on the
C.
striata fillet: close to the head, middle of the fillet and close to the tail. For every body component, the average of two measurements was taken. The L*C*H colour model was then used to translate measurements taken using the L*a*b* colour model, where L* stands for brightness and C* for chroma, which is equal to [(a*)2 + (b*)2]1/2
(Choubert et al., 1997). The same procedure was also followed to analyze the colour of the whole
C.
striata fish before making it into a fillet. The colour was recorded above the lateral line at three points for the entire
C.
striata fish: close to the head, in the middle and close to the tail. The illumination system was rotated 90 degrees between duplicate measurements for each position of the whole fish. For every body component, an average of two measurements was taken to measure the whiteness of the entire fish. The whiteness value of
C.
striata external colour of fish, raw fillets and cooked fillets was obtained using the formula,