Volume 53, Issue 15 pp. 5270-5286

ORIGINAL ARTICLE

Full Access

Dietary lipid requirement of juvenile white-leg shrimp, Penaeus vannamei (Boone, 1931) reared in inland ground saline water of 15 ppt

Prasanta Jana,

Prasanta Jana

orcid.org/0000-0002-2036-1044

Fish Nutrition, Biochemistry and Physiology Division, ICAR - Central Institute of Fisheries Education, Mumbai, India

Search for more papers by this author

Narottam Prasad Sahu,

Corresponding Author

Narottam Prasad Sahu

orcid.org/0000-0002-7100-731X

Fish Nutrition, Biochemistry and Physiology Division, ICAR - Central Institute of Fisheries Education, Mumbai, India

Correspondence

Narottam Prasad Sahu, Fish Nutrition, Biochemistry and Physiology Division, ICAR-Central Institute of Fisheries Education, Versova, Mumbai 400 061, India.

Email: [email protected]; [email protected]

Search for more papers by this author

Parimal Sardar,

Parimal Sardar

orcid.org/0000-0003-0126-2486

Fish Nutrition, Biochemistry and Physiology Division, ICAR - Central Institute of Fisheries Education, Mumbai, India

Search for more papers by this author

Tincy Varghese,

Tincy Varghese

Fish Nutrition, Biochemistry and Physiology Division, ICAR - Central Institute of Fisheries Education, Mumbai, India

Search for more papers by this author

Ashutosh Dharmendra Deo,

Ashutosh Dharmendra Deo

Fish Nutrition, Biochemistry and Physiology Division, ICAR - Central Institute of Fisheries Education, Mumbai, India

Search for more papers by this author

Nazeemashahul Shamna,

Nazeemashahul Shamna

Fish Nutrition, Biochemistry and Physiology Division, ICAR - Central Institute of Fisheries Education, Mumbai, India

Search for more papers by this author

Vungurala Harikrishna,

Vungurala Harikrishna

ICAR-Central Institute of Fisheries Education, Rohtak, India

Search for more papers by this author

Mritunjoy Paul,

Mritunjoy Paul

Fish Nutrition, Biochemistry and Physiology Division, ICAR - Central Institute of Fisheries Education, Mumbai, India

Search for more papers by this author

Nisha Chuphal,

Nisha Chuphal

Fish Nutrition, Biochemistry and Physiology Division, ICAR - Central Institute of Fisheries Education, Mumbai, India

Search for more papers by this author

Gopal Krishna,

Gopal Krishna

Fish Genetics and Biotechnology Division, ICAR - Central Institute of Fisheries Education, Mumbai, India

Search for more papers by this author

Prasanta Jana,

Prasanta Jana

orcid.org/0000-0002-2036-1044

Fish Nutrition, Biochemistry and Physiology Division, ICAR - Central Institute of Fisheries Education, Mumbai, India

Search for more papers by this author

Narottam Prasad Sahu,

Corresponding Author

Narottam Prasad Sahu

orcid.org/0000-0002-7100-731X

Fish Nutrition, Biochemistry and Physiology Division, ICAR - Central Institute of Fisheries Education, Mumbai, India

Correspondence

Narottam Prasad Sahu, Fish Nutrition, Biochemistry and Physiology Division, ICAR-Central Institute of Fisheries Education, Versova, Mumbai 400 061, India.

Email: [email protected]; [email protected]

Search for more papers by this author

Parimal Sardar,

Parimal Sardar

orcid.org/0000-0003-0126-2486

Fish Nutrition, Biochemistry and Physiology Division, ICAR - Central Institute of Fisheries Education, Mumbai, India

Search for more papers by this author

Tincy Varghese,

Tincy Varghese

Fish Nutrition, Biochemistry and Physiology Division, ICAR - Central Institute of Fisheries Education, Mumbai, India

Search for more papers by this author

Ashutosh Dharmendra Deo,

Ashutosh Dharmendra Deo

Fish Nutrition, Biochemistry and Physiology Division, ICAR - Central Institute of Fisheries Education, Mumbai, India

Search for more papers by this author

Nazeemashahul Shamna,

Nazeemashahul Shamna

Fish Nutrition, Biochemistry and Physiology Division, ICAR - Central Institute of Fisheries Education, Mumbai, India

Search for more papers by this author

Vungurala Harikrishna,

Vungurala Harikrishna

ICAR-Central Institute of Fisheries Education, Rohtak, India

Search for more papers by this author

Mritunjoy Paul,

Mritunjoy Paul

Fish Nutrition, Biochemistry and Physiology Division, ICAR - Central Institute of Fisheries Education, Mumbai, India

Search for more papers by this author

Nisha Chuphal,

Nisha Chuphal

Fish Nutrition, Biochemistry and Physiology Division, ICAR - Central Institute of Fisheries Education, Mumbai, India

Search for more papers by this author

Gopal Krishna,

Gopal Krishna

Fish Genetics and Biotechnology Division, ICAR - Central Institute of Fisheries Education, Mumbai, India

Search for more papers by this author

First published: 19 July 2022

https://doi.org/10.1111/are.16012

Citations: 2

Share a link

Email
Wechat
Bluesky

Abstract

A 60-day feeding trial was conducted to evaluate the optimum dietary lipid level in terms of growth performance, digestive enzymes and hemato-biochemical parameters of white-leg shrimp, Penaeus vannamei juveniles in inland ground saline water (IGSW) of 15 ppt salinity. The shrimps (avg. wt., 4.04 ± 0.03 g) were randomly distributed in six groups in triplicates (15 shrimps/tank, 290 L), viz., TCL40 (40 g/kg lipid)_, TCL60 (60 g/kg lipid)_, TCL80 (80 g/kg lipid), TCL100 (100 g/kg lipid), TCL120 (120 g/kg lipid) and TCL140 (140 g/kg lipid). Six semi-purified hetero-lipidic (40-140 g lipid/kg), hetero-caloric (376–426 Kcal DE/100 g) and iso-nitrogenous (400 g crude protein/kg) diets were prepared and fed to respective groups four times daily. Weight gain (WG), specific growth rate (SGR), feed conversion ratio (FCR), and protein efficiency ratio (PER) showed higher quadratic relations (R² = 0.97, 0.98, 0.94 and 0.87, respectively) to dietary lipid levels (p < 0.05) with enhancing dietary lipid up to 60 g/kg. Whole body moisture and lipid contents exhibited a high linear (R² = 0.87 and 0.98) and quadratic (R² = 0.88 and 0.98) (p < 0.05) inverse relationship with enhancing dietary lipid levels up to 140 g/kg. Hemolymph hemocyanin, serum total protein and glucose level did not differ significantly (p > 0.05), but the TCL60 group showed significantly (p < 0.05) higher serum cholesterol and triglyceride levels. Serum osmolality, osmoregulatory capacity, and branchial Na⁺/K⁺-ATPase activity was similar (p > 0.05) among the groups. Hepatopancreatic lipase and amylase activity significantly (p < 0.05) increased, but trypsin activity did not vary significantly (p > 0.05) among the groups. Second-degree polynomial analysis with WG, SGR and FCR indicated that a 55.80–58.80 g lipid/kg diet is optimum for P. vannamei juveniles in IGSW of 15 ppt salinity.

1 INTRODUCTION

The fastest-growing aquaculture industry continues to grow among all the agri and allied sectors and contributes a substantial share of the global food basket to alleviate global hunger (FAO, 2020). The stagnation of marine capture fisheries drives the demand for fish from the culture-based fisheries sector to an increased extent (FAO, 2018). Further, horizontal expansion of land-based aquaculture creates a major conflict with the growing demand for food and land resources for the human population. Therefore, by 2030, the huge targeted fish production can only be achieved using new or alternative aqua farming practices, i.e., adopting unutilized or underutilized natural resources for commercial aqua farming. More than 385 million hectares of land are salt-affected and are expected to rise by 50% by 2050 (Jamil et al., 2011; Partridge et al., 2008). These vast natural resources are not suitable for agricultural activities (Bui et al., 2017). Exploring the degraded soils and under-utilized inland ground saline water (IGSW, salinity 3–25 ppt in India) for aquaculture practices can provide employment opportunities and livelihood generation through enhanced fish and shrimp production (Talukdar et al., 2021). However, in contrast to brackish water and sea water, ionic imbalance (variable concentration of Mg²⁺ with high Ca²⁺ and low K⁺ ions) (Aklakur, 2017) of IGSW can cause a pronounced effect on the growth and physiological homeostasis of the cultured animals (Antony et al., 2020; Saoud et al., 2003; Singha et al., 2020; Talukdar et al., 2020). Available calcium to magnesium ratio (Ca: Mg) in IGSW is higher than in brackish water and sea water, resulting in poor growth performance and less farmed shrimp production (Talukdar et al., 2021). Besides, K⁺ is essential for osmoregulatory adaptations by activation of Na⁺/K⁺-ATPase in the body of shrimp (Roy et al., 2007). Therefore, potassium supplementation through diet and water fortification can be a practical approach for successful shrimp culture in inland ground water of different ambient salinities (Antony et al., 2015; Davis et al., 2005; Jahan et al., 2018; Raizada et al., 2015; Saoud et al., 2007).

Throughout the globe, the farming of high prized euryhaline crustacean, Pacific white-leg shrimp, Penaeus vannamei is very popular due to its faster growth rate, higher disease resistance and better survival with promising economic return (Cuzon et al., 2004; Flores et al., 2007; Hamidoghli et al., 2018). Moreover, comparatively lower dietary crude protein requirements than black tiger shrimp, Penaeus monodon and tolerance of widespread salinity (1–50 ppt, recommended salinity 15–30 ppt) make P. vannamei more suitable species for IGSW (Lightner et al., 2009; Samocha et al., 2002; Yun et al., 2016; Zhou et al., 2014). Huong et al. (2010) and Li et al. (2017) reported that P. vannamei could adjust ambient salinity changes through osmoregulation comprising amino acid balance pools, regulating the volume of cells and activation of enzymes of ion transporter system. However, drastic change in water salinity greatly influences the growth performance, survival rate, physio-metabolic responses and finally, resistance to diseases, resulting in the diversion of a higher amount of energy to compensate the metabolic needs (Diaz et al., 2001; Tseng & Hwang, 2008; Wang et al., 2014). Thus, nutritional intervention through diet could be a cost-effective and useful practice to uphold the basic physio-metabolic responses for improving the growth performance of shrimp in IGSW (Li et al., 2008; Ur-Rahman et al., 2005; Wang et al., 2015; Xu et al., 2016).

Dietary lipid is a concentrated source of digestible energy. It serves as a basic precursor of EFAs (essential fatty acids), phospholipids, glycolipids and sterols mostly required for the growth, physio-metabolic and immune responses of shrimp (González-Félix, Gatlin III, et al., 2002; González-Félix, Lawrence, et al., 2002; Xie et al., 2019; Zhang et al., 2013). Besides, it also carries essential fat-soluble vitamins (He et al., 1992), maintaining cellular and sub cellular membrane structure, function and integrity (Bou et al., 2017), synthesis of moulting hormones (Arts & Kohler, 2009; González-Félix et al., 2010) and osmoregulatory adaptations (Jannathulla et al., 2019). Dietary lipid also has the protein-sparing effect of reducing the nitrogenous wastes from protein catabolism in the rearing water (Luo et al., 2005).

Several authors have investigated and reported that the dietary lipid requirements of P. vannamei ranged between 50 and140 g/kg (Goda, 2008; González-Félix, Lawrence, et al., 2002; Tzeng et al., 2004). Dietary lipid requirement changes with the salinity, different life stages and the culture system (Chen et al., 2014; González-Félix, Lawrence, et al., 2002; Hamidoghli et al., 2020; Jannathulla et al., 2019; Toledo et al., 2016; Xie et al., 2019; Zhu et al., 2010). Xu et al. (2018) reported that 90 g/kg dietary lipid is optimum for P. vannamei juveniles regardless of salinity (3 and 25 ppt) in terms of growth performance and nutrient utilization, but feeding higher lipid levels at 120 g/kg adversely affects the growth and oxidative damage of the hepatopancreas of the animal. Jannathulla et al. (2019) described higher growth and better survival of shrimp P. vannamei due to feeding of 60 g lipid/kg with12 g/kg essential fatty acids at 25 ppt ambient salinity. However, Xie et al. (2019) described relatively higher dietary lipid requirement (118–124 g lipid/kg diet) of white-leg shrimp post-larvae to improve the growth, survivability, immune response and stress resistance at 28 ppt water salinity. Similarly, Zhang et al. (2013) recommended that a 100–120 g lipid/kg dietary level is optimum for P. vannamei juveniles in terms of growth and immunity reared at 7 ppt salinity. Hamidoghli et al. (2020) and Toledo et al. (2016) reported that P. vannamei juveniles require 56–60 g/kg and 8.5–9.5 g/kg dietary lipids, respectively, in biofloc based culture system. However, earlier studies (González-Félix, Gatlin III, et al., 2002; González-Félix, Lawrence, et al., 2002) demonstrated that dietary 30–90 g lipid/kg is optimum for better growth and survival of white-leg shrimp juveniles.

With this backdrop, the current experiment was designed and aimed to evaluate the dietary lipid requirement based on the growth performance, nutrient utilization, digestive enzymes and hemato-biochemical parameters of juvenile white-leg shrimp, P. vannamei, cultured in 15 ppt IGSW. This is the first report of its kind for white-leg shrimp reared in IGSW. We also optimized the protein requirement for the P. vannamei diet specific to IGSW (Jana et al., 2021). The outcome of the current study will serve as the baseline information for the global aqua feed industry for precise shrimp feed formulation, especially for the vibrant commercial inland saline water shrimp culture.

2 MATERIALS AND METHODS

2.1 Ethical declarations

The current study strictly adhered to animal welfare norms for scientific investigations after approval from the Animal Ethics Committee of ICAR-Central Institute of Fisheries Education (CIFE), India and CPCSEA (Committee for the Purpose of Control and Supervision of Experiments on Animals), Ministry of Environment & Forests (Animal Welfare Division), Government of India.

2.2 Lab animal

White-leg shrimp juveniles (one thousand fifteen hundred) were carefully packed in polythene triple-layered bags containing well-oxygenated 15 ppt IGSW and immediately transported from Anwal Shrimp farm, Rohtak, Haryana, to the Experimental Laboratory for Shrimp of ICAR-CIFE, Rohtak centre. For acclimatization, shrimps were immediately shifted to circular tanks (10.03 m² × 0.87 m, 1000 L water capacity, 800 L water volume) filled with IGSW of 15 ppt under natural photoperiod with vigorous aeration. During 15 days acclimatization period, shrimps were fed four times a day with a commercial diet containing 400 g/kg crude protein and 50 g/kg dietary lipid.

2.3 Formulation and preparation of diets

Six iso-nitrogenous (400 g/kg dietary protein), hetero-lipidic (40 to 140 g/kg dietary lipid), and hetero-energetic (376 to 426 Kcal DE/100 g) semi-purified experimental diets were formulated (Table 1) with varying lipid levels viz., TCL40 (40 g/kg lipid), TCL60 (60 g/kg lipid), TCL80 (80 g/kg lipid), TCL100 (100 g/kg lipid), TCL120 (120 g/kg lipid) and TCL140 (140 g/kg lipid). All the finely milled ingredients were properly weighed according to the formulation and kept in separate plastic airtight containers. Except for oils and additives, all the ingredients were uniformly mixed in a mechanical mixer. The dough was prepared and steam cooked for 25 min. After cooling at room temperature, additives and oils were homogenously mixed with the dough and pellets (2 mm dia) were prepared using a pelletizer (S.B. Panchual & Co. Ltd, India). The feed pellets were air-dried at room temperature afterwards in a hot air oven at 42°C till to achieve around 10% moisture level. The dried pellets were properly packed, sealed, labelled, and kept at 4°C until further use.

TABLE 1. Formulation and proximate composition of the experimental diets

	TCL40	TCL60	TCL80	TCL100	TCL120	TCL140
	Diets (experimental groups)^a
Ingredients composition (g/kg)
Fish Meal	200	200	200	200	200	200
Soybean meal	100	100	100	100	100	100
Casein	208	208	208	208	208	208
Gelatin	52	52	52	52	52	52
Starch	202.5	182.5	162.5	142.5	122.5	102.5
Dextrin	150	150	150	150	150	150
Cellulose	19.4	19.4	19.4	19.4	19.4	19.4
Fish oil	15	35	55	75	95	115
CMC	25	25	25	25	25	25
Vit. min. mix^b	15	15	15	15	15	15
Soy lecithin	5	5	5	5	5	5
Cholesterol	2	2	2	2	2	2
BHT	0.2	0.2	0.2	0.2	0.2	0.2
Betaine hydrochloride	5	5	5	5	5	5
Choline chloride	0.3	0.3	0.3	0.3	0.3	0.3
Stay C 35^c	0.5	0.5	0.5	0.5	0.5	0.5
Vit. E	0.1	0.1	0.1	0.1	0.1	0.1
Total	1000	1000	1000	1000	1000	1000
Proximate composition (g/kg dry matter basis)
Dry matter	911.2	912.6	909.8	910.5	913.2	911.6
Crude protein	401.5	400.9	401.0	401.3	400.3	401.7
Ether extract	40.8	60.5	81.3	102.1	121.5	141.9
Crude fibre	29.2	29.1	29.4	29.5	29.6	30.1
Nitrogen free extract	447.0	427.7	407.7	384.6	366.9	345.4
Total ash	81.4	81.6	80.5	82.3	81.7	80.9
GE (Kcal/100 g)	449.98	460.03	471.36	481.44	491.72	502.84
DE^d (Kcal/100 g)	376.11	385.86	396.58	405.87	416.10	426.57
P/E^e (mg CP/Kcal DE)	106.75	103.90	101.11	98.87	96.20	94.17

Note: Proximate composition is expressed as mean (n = 3).
Abbreviations: BHT, butylated hydroxytoluene; CMC, carboxymethyl cellulose; GE, gross energy.
^a TCL40 (40 g/kg dietary lipid), TCL60 (60 g/kg dietary lipid), TCL80 (80 g/kg dietary lipid), TCL100 (100 g/kg dietary lipid), TCL120 (120 g/kg dietary lipid), TCL140 (140 g/kg dietary lipid).
^b Composition of vitamin-mineral mix (PRE-EMIX PLUS) (quantity/kg): Vitamin A, 5,500,000 IU; Vitamin D3, 1,100,000 IU; Vitamin B2, 2000 mg; Vitamin E, 750 mg; Vitamin K, 1000 mg; Vitamin B6, 1000 mg; Vitamin B12, 6 mg; Calcium Pantothenate, 2500 mg; Nicotinamide, 10 g; Choline Chloride, 150 g; Mn, 27,000 mg; I, 1000 mg; Fe, 7500 mg; Zn, 5000 mg; Cu, 2000 mg; Co, 450 L-lysine, 10 g; dl- Methionine, 10 g; Selenium 125 mg; Vitamin C,2500 mg.
^c Stay C 35, protected vitamin C.
^d DE, digestible energy (Kcal/100 g) = {4 × CP (g/100 g) + 9 × EE (g/100 g) + 4 × NFE (g/100 g)} (Halver, 1976).
^e P/E, protein to energy ratio (mg CP/Kcal DE) = {(CP% × 1000)/DE}.

2.4 Collection, filling and storage of IGSW

IGSW of identical salinity was taken out and passed through a fixed filter bag (80 μm mesh size). Subsequently, the filtered water was filled into rectangular tanks (3.1 × 2.1 × 1.1 m³, 10,000 L capacity) and kept for a week. After that, IGSW was pumped into circular FRP storage tanks (10.03 m² × 0.87 m, 1000 L water capacity). These storage tanks were fitted with round a clock aeration facility and fortified with commercially available muriate of potash (KCl) as and when required. The requisite was calculated using the formula: K⁺ requirement for fortification = {(Experimental salinity level × 10.7) − IGSW available K⁺} (Davis et al., 2005).

2.5 Setting up of experiment and feeding trial

For setting up of the trial, 18 circular tanks (8.82 m² × 51 m, 350 L capacity and 290 L water volume) were thoroughly washed with potassium permanganate (KMnO₄) solution (5 g/L) followed using freshwater and dried under direct sunlight. The tanks were then filled with fortified IGSW with continuous aeration. Before stocking, shrimps were starved overnight, and a digital weighing balance recorded carefully initial weight. Well acclimatized two hundred and seventy shrimps (avg. wt., 4.04 ± 0.03 g) were randomly distributed (15 shrimps/tank) among six experimental groups in triplicates following a completely randomized design (CRD). Respective diets were used to feed the shrimp four times a day (7:30, 13:30, 18:30 and 23:30 h). Faecal matter from each tank was siphoned out prior to next day's feeding, and an equal volume of water was replenished from the storage tank. About 20–25% water from each tank was exchanged with IGSW of 15 ppt salinity from the storage facility at the interval of 3 days during the experimental period. Fortnightly, the total biomass of the shrimps from each tank was taken to calculate the daily ration size. Mortality of shrimps was recorded to calculate the survival percentage.

Water quality parameters like temperature, pH and dissolved oxygen (DO) were estimated using a digital thermometer (Oxyguard International, India), digital pH meter (Manti Lab India, Pvt. Ltd.) and portable DO meter (Manti Lab India, Pvt. Ltd.), respectively, thrice a day. At every 3 days intervals, Physico-chemical parameters of water like salinity, free carbon-di-oxide (CO₂)_, total alkalinity, total hardness, ammonia-N (TA-N), nitrite-N (NO₃-N), nitrate-N (NO₂-N), calcium, magnesium were analysed as per the method described using APHA (1998). A digital automated flame photometer (1835, Esico India Pvt. Ltd.) was used to determine K⁺ concentration in IGSW. Experimental IGSW quality parameters like temperature ranged between 26.03 ± 0.3 to 27.15 ± 0.56°C, pH 8.35 ± 0.11 to 8.47 ± 0.18, salinity 15.15 ± 0.18 to 15.36 ± 0.17 g/L, dissolved oxygen 6.81 ± 0.42 to 7.09 ± 0.30 mg/L, total alkalinity 297.55 ± 6.71 to 309.59 ± 6.52 mg/L, total hardness 2948.00 ± 9.61 to 2975.00 ± 17.73 mg/L with no detectable free CO₂. The TA-N, NO₂-N and NO₃-N were found in the range of 0.17 ± 0.03 to 0.22 ± 0.03 mg/L, 0.12 ± 0.01 to 0.14 ± 0.01 mg/L, 0.24 ± 0.02 to 0.30 ± 0.03 mg/L, respectively, during the experimental period. The concentrations of calcium, magnesium and potassium were ranged from 241.34 ± 3.97 to 250.98 ± 3.61 mg/L, 533.79 ± 3.02 to 541.72 ± 3.46 mg/L, and 100.53 ± 0.67 to 101.46 ± 0.93 mg/L, respectively (Table 2).

TABLE 2. Water quality parameters of different experimental groups during experimental period of 60 days

Parameters	^aDiets (experimental groups)
Parameters	TCL40	TCL60	TCL80	TCL100	TCL120	TCL140	p-value
Temperature (°C)	26.17 ± 0.46	26.03 ± 0.34	26.79 ± 0.26	27.15 ± 0.56	26.59 ± 0.23	27.07 ± 0.21	0.761
pH	8.44 ± 0.15	8.41 ± 0.13	8.47 ± 0.18	8.36 ± 0.09	8.41 ± 0.17	8.35 ± 0.11	0.653
Salinity (g/L)	15.15 ± 0.18	15.21 ± 0.23	15.27 ± 0.09	15.31 ± 0.19	15.36 ± 0.17	15.34 ± 0.12	0.723
DO (mg/L)	6.81 ± 0.42	6.92 ± 0.46	6.99 ± 0.24	6.94 ± 0.43	7.05 ± 0.52	7.09 ± 0.30	0.638
Free CO₂ (mg/L)	Nil	Nil	Nil	Nil	Nil	Nil	-
Alkalinity (mg/L)	299.74 ± 3.89	297.55 ± 6.71	305.98 ± 4.65	302.36 ± 4.21	305.39 ± 5.46	309.59 ± 6.52	0.823
Hardness (mg/L)	2959.00 ± 14.64	2966.00 ± 15.62	2951.50 ± 11.27	2975.00 ± 17.73	2948.00 ± 9.61	2956.50 ± 10.12	0.757
TA-N (mg/L)	0.22 ± 0.03	0.19 ± 0.04	0.20 ± 0.03	0.19 ± 0.04	0.17 ± 0.03	0.18 ± 0.02	0.569
NO₂-N (mg/L)	0.14 ± 0.01	0.13 ± 0.02	0.14 ± 0.01	0.12 ± 0.01	0.12 ± 0.02	0.13 ± 0.03	0.618
NO₃-N (mg/L)	0.30 ± 0.03	0.28 ± 0.02	0.29 ± 0.04	0.24 ± 0.02	0.27 ± 0.02	0.28 ± 0.03	0.702
Ca²⁺ (mg/L)	241.34 ± 3.97	243.86 ± 2.68	247.80 ± 3.42	248.67 ± 2.73	250.98 ± 3.61	242.05 ± 4.74	0.839
Mg²⁺ (mg/L)	533.79 ± 3.02	535.62 ± 2.95	538.74 ± 2.79	539.78 ± 3.99	541.72 ± 3.46	534.09 ± 2.16	0.694
K⁺ (mg/L)	100.61 ± 1.24	100.53 ± 0.67	100.58 ± 0.48	101.21 ± 0.74	101.46 ± 0.93	101.01 ± 1.11	0.688

Notes: Data are expressed as mean ± SE (n = 6); Mean values in the same row with different superscripts differ significantly (p < 0.05).
Abbreviations: CO_2, carbon di-oxide; DO, dissolved oxygen; NO₂-N, nitrite nitrogen; NO₃-N, nitrate nitrogen; TA-N, total ammonia nitrogen.
^a TCL40-TCL140, 40–140 g/kg dietary crude lipid.

2.6 Sampling

Prior to commencement of the experiment, 25 shrimps were randomly collected for initial body composition analysis. After completion of the trial, shrimps were starved overnight to measure final body weight. Randomly three shrimps from each replicate (nine shrimps from a treatment) were taken out for the final proximate composition analysis. Five shrimps from each tank were anaesthetized with chilled iced water and hemolymph was drawn from the shrimp's heart and pooled (Antony et al., 2015). For collection of serum, hemolymph samples were kept in a freezer (4°C) for 20 min followed using crushing and centrifugation at 3500g for 12 min (Tantulo & Fotedar, 2006). Again, from three shrimps of each treatment (one from each replicate), hemolymph was collected and mixed with ice-cold SIC-EDTA anticoagulant in a 1:2 ratio for estimation of hemocyanin content (Vargas-Albores et al., 1993). From these eight shrimps carefully hepatopancreas (weight was recorded for calculation of HPSI) and intestine were dissected out and quickly homogenized (5% homogenate) in iced phosphate buffer solution (pH 7.4) for assay of digestive enzymes. Gills were homogenized (10% homogenate) in iced SEI buffer (pH 7.3) using an automated homogenizer (Miccra, D9, Remi India) under iced conditions (Jana et al., 2021; Saraswathy et al., 2021) for Na⁺/K⁺-ATPase assay. The tissue homogenates were then centrifuged at 7000g for 15 min in a refrigerated centrifuge, and the aliquot supernatant was collected in sample vials. All the samples were properly labelled and stored at −40°C until further use.

2.7 Proximate composition of the diets and tissues

Shrimp whole-body tissue and experimental diets proximate analysis were performed by following AOAC (1995). For moisture analysis, the initial fish sample, final fish samples and diets were oven-dried at 80°C until achieving a constant weight. The micro-Kjeldahl method (Kjelteck, Pelican, India) was followed to estimate the crude protein (CP), but lipid was estimated using solvent extraction using Soxhlet's apparatus (SOCSplus, Pelican, India). Samples were incinerated at 550°C for 5 h in a muffle furnace (WIC, CL Teltow, Australia) for determining the total ash (TA) content. Fat-free samples' crude fibre (CF) content was performed through acid digestion followed using alkaline digestion in fibretec (Tulin Equipments, India). Then the digested sample was incinerated in muffle furnace at 550°C for 5 h. The subtraction method was employed for the calculation of the NFE (nitrogen-free extract) content of the experimental diets as follows:

NFE (g / kg) = \{1000 - (g CP / kg + g EE / kg + g CF / kg + g TA / kg)\}

Gross energy (Kcal/100 g) of the experimental diets' was analysed using a bomb calorimeter (5AECPL, Changhsha Manufacturers Pvt. Ltd. China).

The lipid levels of the experimental diets viz., TCL40, TCL60, TCL80, TCL100, TCL120 and TCL140 were 40.8, 60.5, 81.3, 102.1, 121.5 and 141.9 g/kg, respectively (Table 1). Dietary crude protein (CP), crude fibre (CF), total ash (TA) and nitrogen-free extract (NFE) ranges between 400.3 to 401.7, 29.1 to 30.1, 80.5 to 82.3 and 345.4 to 447 g/kg, respectively, in different experimental diets. The values of gross energy (GE), digestible energy (DE) and protein to energy ratio (P: E) range between 449.98 to 502.84 Kcal/100 g, 376.11 to 426.57 Kcal/100 g and 94.17 to 106.75 mg CP/Kcal DE, respectively (Table 1). Thus, the semi-purified experimental diets were iso-nitrogenous, hetero-lipidic and hetero-energetic.

2.8 Growth metrics, nutrient utilization and survival

The following formulas were used for calculation of growth metrics, nutrient utilization and survival rate of the experimental shrimp:

WG (g) = \{Final wet weight (g) - Initial wetweight in (g)\}

SGR (% / day) = \frac{\{\ln final wet weight (g) - \ln initial wet weight (g)\}}{Feeding days} \times 100

FCR = \frac{\{Feed given (dry weight in g)\}}{\{Live wet weight gain (g)\}}

PER = \frac{\{Live wet weight gain (g)\}}{\{Protein consumed (dry weight in g)\}}

Survival (%) = \frac{Total number of alive shrimps}{Total number of stocked shrimps} \times 100

HPSI (%) = \frac{\{Wet weight of hepatopancreas (g)\}}{\{Wet weight of shrimp weight in (g)\}} \times 100

2.9 Hemato-biochemical analysis

Hemolymph hemocyanin was estimated according to Chen and Cheng (1993a) and Chen and Cheng (1993b). In a quartz cuvette, 10 μl hemolymph was mixed with 990 μl demineralized water and reading was taken in a double beam spectrophotometer (Shimadzu 1800, Shimadzu Corporation, Pvt. Ltd., China) at an optical density of 335 nm. For calculation of the final concentration of hemocyanin, an extinction coefficient of E = 17.26 was used based on the 74 KDa functional subunit of shrimp.

Serum and water osmolality (mOsmol/Kg) was determined in an osmometer (Osmostat035, GMBH Corporation, Germany). Osmoregulatory capacity (OC) (mOsmol/Kg) was calculated using the formula (Lignot et al., 2000):

\begin{matrix} OC (mOsmol / kg) = \\ mean serum osmolality of shrimp - mean water osmolality \end{matrix}

Commercial test kits were procured from Transasia Biotech Pvt. Ltd., ERBA Diagnostics, Solan, India, to estimate serum glucose, total protein, triglyceride and cholesterol.

2.10 Enzyme assay

2.10.1 Quantification of tissue protein

Protein quantification in different tissue homogenates was performed according to Lowry et al. (1951). The values of tissue protein obtained were used for calculating the activities of tissue specific-enzymes.

2.10.2 Activities of Na⁺/K⁺-ATPase

Na⁺/K⁺-ATPase specific activities (micromoles of ADP released/h/mg protein at 37°C) in the gill lamellae were measured according to the method of McCormick (1993).

2.10.3 Activities of digestive enzymes

Trypsin specific activities (units/min/mg protein at 37°C) were estimated using following Na-benzoyl-DL-arginine-p-nitroanilide (BAPNA) substrate method (Garcia-Carreno et al., 1994). The activities of amylases (micromole of maltose released/ mg protein/ min at 37°C) were determined as described by Rick and Stegbauer (1974). Lipase specific activities (units/min/mg protein at 37°C) were assayed according to Cherry and Crandall Jr. (1932).

2.11 Statistical analysis

Results were analysed through one-way analysis of variance (ANOVA) using SPSS (22 for windows) software. Overall, linear and quadratic trends of the parameters dietary lipid levels were confirmed using employing the orthogonal-polynomial contrast analysis. Duncan multiple range tests (DMRT) along with post-hoc analysis were applied to observe the significant differences among the means between the treatments at a 5% probability level (p < 0.05). Second-degree polynomial analysis was executed to optimize the dietary lipid requirement of white-leg shrimp, P. vannamei reared in IGSW of 15 ppt salinity.

3 RESULTS

3.1 Growth metrics, nutrient utilization and survivability of shrimp

Growth metrics and nutrient utilization parameters of juvenile white-leg shrimp, P. vannamei varied significantly (p < 0.05) owing to varying dietary lipid levels (Table 3). The highest (p < 0.05) weight gain (WG) and specific growth rate (SGR) were observed in the TCL60 group, followed by TCL80 and TCL40 group, whereas the TCL140 group exhibited the lowest (p < 0.05) value. Similarly, TCL60 and TCL140 groups exhibited the lowest and highest (p < 0.05) feed conversion ratio (FCR), respectively. However, a decreasing trend was observed for protein efficiency ratio (PER) with the increased lipid levels in the diet. Hepatopancreas-somatic index (HPSI) and survival rate of shrimp were found to be similar (p > 0.05) among the treatment groups. Regression analysis revealed variable linear and quadratic correlation of WG (linear R² = 0.80 and quadratic R² = 0.97), SGR (linear R² = 0.79 and quadratic R² = 0.98) and FCR (linear R² = 0.77 and quadratic R² = 0.94), PER (linear R² = 0.73 and quadratic R² = 0.87) in relation to the dietary lipid levels. However, dietary lipid levels exhibited poor linear and quadratic relations with survival rate (linear R² = 0.09 and quadratic R² = 0.11) and HPSI (linear R² = 0.09 and quadratic R² = 0.11) of the treatment groups. Based on second-degree polynomial analysis of WG, SGR and FCR with the dietary lipid levels, optimum values of dietary lipid were found to be 58.80, 57.0 and 55.80 g/kg, respectively, for white-leg shrimp, P. vannamei reared in IGSW (Figures 1-3).

TABLE 3. Growth performance, nutrient utilization, survivability and hepatopancreatic-somatic index (HPSI) of Penaeus vannamei juveniles fed diets with varying levels of dietary lipid reared in IGSW of 15 ppt for 60 days

Diets (experimental groups)^a		Parameters				Survival (%)	HPSI (%)
Diets (experimental groups)^a		WG (g)	SGR (%/day)	FCR	PER	Survival (%)	HPSI (%)
TCL40		10.41^d	2.12^d	1.45^c	1.72^d	91.11	1.76
TCL60		11.11^f	2.20^f	1.27^a	1.97^f	93.33	1.72
TCL80		10.66^e	2.15^e	1.41^b	1.77^e	84.45	1.73
TCL100		9.68^c	2.03^c	1.58^d	1.58^c	88.89	1.72
TCL120		8.53^b	1.89^b	1.75^e	1.43^b	88.89	1.73
TCL140		7.31^a	1.73^a	1.96^f	1.27^a	86.67	1.71
SEM		0.32	0.04	0.06	0.06	1.20	0.01
Contrast analysis, p value
Overall		<0.001	<0.001	<0.001	<0.001	0.366	0.692
Linear		<0.001	<0.001	<0.001	<0.001	0.219	0.275
Quadratic		<0.001	<0.001	<0.001	<0.001	0.622	0.572
Regression equation, R² value
Linear	Equation^b	y = −0.3464x + 12.041	y = −0.0431x + 2.3227	y = 0.0591x + 1.1547	y = −0.0575x + 2.0256	y = −0.4444x + 92	y = −0.0029x + 1.7464
Linear	R²	0.80	0.79	0.77	0.73	0.09	0.09
Quadratic	Equation^b	y = −0.0553x² + 0.4277x + 9.9769	y = −0.0072x² + 0.0572x + 2.0554	y = 0.0094x²–0.0726x + 1.5059	y = −0.0084x² + 0.0605x + 1.7109	y = 0.0595x²–1.2778x + 94.222	y = 0.0005x2–0.01x + 1.7653
Quadratic	R²	0.97	0.98	0.94	0.87	0.11	0.11

Notes: Data are expressed as mean (n = 3); Mean values in the same column with different superscripts differ significantly (p < 0.05).
Abbreviations: FCR, feed conversion ratio; HPSI, hepatopancreatic-somatic index; PER, protein efficiency ratio; SEM, average standard error of means; SGR, specific growth rate; WG, weight gain.
^a TCL40-TCL140, 40–140 g/kg dietary crude lipid.
^b In equation, ‘x’ and ‘y’ represents dietary lipid levels and the respective parameters.

Details are in the caption following the image — **FIGURE 1**
Open in figure viewer PowerPoint

Second-order polynomial regression analysis between weight gain (g) and dietary lipid levels (g/kg) for *Penaeus vannamei* juveniles reared in inland ground saline water of 15 ppt for 60 days.

3.2 Proximate composition of the whole body of shrimp

Proximate analysis of whole shrimp body showed that moisture and lipid contents exhibited a significant (p < 0.05) inverse relationship with the increasing dietary lipid levels (Table 4). The lowest and highest (p < 0.05) moisture and lipid contents in shrimp's whole body were observed in TCL140_, TCL40 and TCL40_, TCL140 groups, respectively. However, whole shrimp body protein and ash content did not vary significantly (p > 0.05) among the treatment groups with the increasing levels of dietary lipid. Regression analysis of shrimp whole body moisture (linear R² = 0.87 and quadratic R² = 0.88) and crude lipid (linear R² = 0.98 and quadratic R² = 0.98) showed a linear and quadratic relation, whereas shrimp whole body protein and total ash showed no defined correlation with respect to crude lipid level of the experimental diets.

TABLE 4. Whole body proximate composition (g/kg wet weight basis) of Penaeus vannamei juveniles fed diets with varying levels of dietary lipid reared in IGSW of 15 ppt for 60 days

Diets (experimental groups)^a		Proximate composition (g/kg)
Diets (experimental groups)^a		Moisture	Crude protein	Crude lipid	Total ash
IBC		748.67^d	175.72	12.55^a	32.14
FBC
TCL40		748.50^d	175.83	12.60^a	32.01
TCL60		741.73^c	175.10	18.97^b	33.13
TCL80		732.90^b	176.60	37.50^c	32.20
TCL100		733.37^b	175.10	44.67^d	31.90
TCL120		724.63^a	174.73	56.30^e	31.40
TCL140		720.03^a	175.53	63.20^f	31.93
SEM		1.88	0.37	4.45	0.52
Contrast analysis, p value
Overall		<0.001	0.808	<0.001	0.976
Linear		<0.001	0.632	<0.001	0.647
Quadratic		0.586	0.983	0.002	0.927
Regression equation, R² value
Linear	Equation^b	y = −2.7594x + 752.84	y = −0.0586x + 175.89	y = 5.3167x + 1.6556	y = −0.0833x + 32.678
Linear	R²	0.87	0.02	0.98	0.02
Quadratic	Equation^b	y = 0.0503x²–3.4634x + 754.72	y = 0.0009x² − 0.0711x + 175.93	y = −0.1113x² + 6.875x - 2.5	y = −0.0057x² − 0.0042x + 32.467
Quadratic	R²	0.88	0.02	0.98	0.02

Notes: Data are expressed as mean (n = 3); Mean values in the same column with different superscripts differ significantly (p < 0.05).
Abbreviations: FBC, final whole body composition; IBC, initial whole body composition; SEM, average standard error of means.
^a TCL40-TCL140, 40–140 g/kg dietary crude lipid.
^b In equation, ‘x’ and ‘y’ represents dietary lipid levels and the respective parameters.

3.3 Hemolymph hemocyanin and serum biochemical indices

Total triglyceride and cholesterol contents of serum significantly (p < 0.05) increased and decreased, respectively, in a similar fashion to the dietary lipid levels (Table 5). However, hemolymph hemocyanin and concentrations of glucose and total protein in the serum did not show any significant (p > 0.05) variations between the groups. On the other hand, there was a significant (p < 0.05) increment in serum triglyceride and cholesterol contents while enhancing the lipid levels in the diet up to the TCL60 group (60 g lipid/kg), and then gradually decreased with the lowest (p < 0.05) value observed in TCL140 (140 g lipid/kg diet) group. Regression analysis of serum total protein, hemocyanin and glucose contents did not show any defined correlation while serum cholesterol (quadratic R² = 0.60) and triglyceride (linear R² = 0.68 and quadratic R² = 0.79) exhibited a high correlation with the dietary lipid levels.

TABLE 5. Hemato-biochemical parameters of Penaeus vannamei juveniles fed diets with varying levels of dietary lipid reared in IGSW of 15 ppt for 60 days

Diets (experimental groups)^a		Total Protein^b (g/dL)	Hemocyanin (Hc)^c (mmol/L)	Glucose^c (mg/dL)	Cholesterol^c (mg/dL)	Triglyceride^c (mg/dL)
TCL40		26.65	1.75	18.45	166.87^a	189.68^c
TCL60		24.61	1.74	20.60	178.93^c	209.45^e
TCL80		25.78	1.73	22.29	173.71^c	199.64^d
TCL100		26.11	1.73	19.78	162.95^b	176.59^b
TCL120		25.88	1.71	21.26	156.62^a	163.15^a
TCL140		26.35	1.75	20.98	151.60^a	158.26^a
SEM		0.26	0.01	0.72	2.57	4.58
Contrast analysis, p value
Overall		0.281	0.910	0.787	<0.001	<0.001
Linear		0.599	0.677	0.463	<0.001	<0.001
Quadratic		0.224	0.389	0.471	<0.001	<0.001
Regression equation, R²value
Linear	Equation^d	y = 0.038x + 25.631	y = −0.0013x + 1.7429	y = 0.173x + 19.348	y = −1.2003x + 171.18	y = −4.558x + 214.7
Linear	R²	0.02	0.01	0.04	0.14	0.68
Quadratic	Equation^d	y = 0.0309x²–0.3949x + 26.785	y = 0.0009x² − 0.0142x + 1.7773	y = −0.0581x² + 0.9869x + 17.178	y = −0.7137x² + 8.7912x + 144.54	y = −0.6151x² + 4.0541x + 191.74
Quadratic	R²	0.10	0.07	0.08	0.60	0.79

Notes: Data are expressed as mean (n = 3); Mean values in the same column with different superscripts differ significantly (p < 0.05).
Abbreviation: SEM, average standard error of means.
^a TCL40-TCL140, 40–140 g/kg dietary crude lipid.
^b Using hemolymph.
^c Using serum.
^d In equation, ‘x’ and ‘y’ represents dietary lipid levels and the respective parameters.

3.4 Na⁺/K⁺-ATPase enzyme activity, osmolality and osmoregulatory capacity (OC) of shrimp

Na⁺/K⁺-ATPase enzyme-specific activity in gill, osmoregulatory capacity and osmolality of cultured water and shrimp serum exhibited no significant (p > 0.05) relationship with respect to different lipid levels of the diet (Table 6). Very poor linear and quadratic correlation of water and shrimp serum osmolality, Na⁺/K⁺-ATPase activities in gill and osmoregulatory capacity were observed in relation to dietary lipid levels.

TABLE 6. Osmolality, osmoregulatory capacity (OC) and branchial Na⁺/K⁺-ATPase activity of Penaeus vannamei juveniles fed diets with varying levels of dietary lipid reared in IGSW of 15 ppt for 60 days

Diets (experimental groups)^a		Serum osmolarity^b	Water osmolarity^b	Osmoregulatory capacity^b (OC)	Na⁺/K⁺-ATPase^c
TCL40		687.32	251.64	435.68	13.67
TCL60		687.45	248.91	438.54	13.17
TCL80		686.72	251.10	435.62	13.85
TCL100		684.62	248.18	436.45	13.38
TCL120		688.12	252.06	436.06	13.06
TCL140		688.33	248.58	439.76	12.96
SEM		1.34	0.94	1.30	0.20
Contrast analysis, p value
Overall		0.984	0.792	0.944	0.827
Linear		0.879	0.682	0.658	0.359
Quadratic		0.63	0.897	0.674	0.692
Regression equation, R² value
Linear	Equation^d	y = 0.0712x + 686.6	y = −0.1254x + 250.95	y = 0.1966x + 435.64	y = −0.0618x + 13.78
Linear	R²	0.00	0.01	0.02	0.07
Quadratic	Equation^d	y = 0.0774x² − 1.0126x + 689.49	y = 0.0135x² − 0.3143x + 251.46	y = 0.0639x² − 0.6982x + 438.03	y = −0.009x² + 0.0642x + 13.444
Quadratic	R²	0.02	0.01	0.03	0.08

Notes: Data are expressed as mean (n = 3); Mean values in the same column with different superscripts differ significantly (p < 0.05).
Abbreviation: SEM, average standard error of means.
^a TCL40-TCL140, 40–140 g/kg dietary crude lipid.
^b Water osmolality, serum osmolality and osmoregulatory capacity (OC) are expressed as mOsmol/kg.
^c Branchial Na⁺/K⁺-ATPase activity is expressed as micromoles of ADP released/hour/mg protein at 37°C.
^d In equation, ‘x’ and ‘y’ represents dietary lipid levels and the respective parameters.

3.5 Hepatopancreatic and intestinal digestive enzymes activities

Hepatopancreatic lipase and amylase specific activities showed significant (p < 0.05) differences among the groups in relation to varying dietary lipid levels, with the highest (p < 0.05) activities were observed inTCL80 (80 g lipid/kg diet) and TCL120 (120 g lipid/kg diet) groups, respectively, and beyond that level, the activity of these enzymes gradually decreased (Table 7). Regression analysis revealed that dietary lipid levels exhibited poor linear and quadratic relations with hepatopancreatic and intestinal trypsin, intestinal amylase and intestinal lipase enzyme activities. However, the activities of hepatopancreatic amylase (linear R² = 0.35 and quadratic R² = 0.51) and lipase (quadratic R² = 0.90) enzymes exhibited variable linear and quadratic relations with graded levels of dietary lipid.

TABLE 7. Digestive enzyme activities in the hepatopancreas and intestine of Penaeus vannamei juveniles fed diets with varying levels of dietary lipid reared in IGSW of 15 ppt for 60 days

Diets (experimental groups)^a		Digestive enzymes
		Trypsin^b		Amylase^c		Lipase^d
		Hepatopancreas	Intestine	Hepatopancreas	Intestine	Hepatopancreas	Intestine
TCL40		9.48	9.08	1.93^ab	1.69	2.36^b	8.67
TCL60		9.55	9.11	1.84^a	1.78	2.57^c	8.69
TCL80		9.54	9.11	2.44^cd	1.66	2.75^d	8.67
TCL100		9.48	9.15	2.56^e	1.75	2.67^cd	8.69
TCL120		9.55	9.08	2.62^ef	1.68	2.44^b	8.68
TCL140		9.54	9.08	2.32^c	1.68	2.21^a	8.71
SEM		0.02	0.02	0.09	0.02	0.05	0.03
Contrast analysis, p value
Overall		0.861	0.980	0.009	0.204	<0.001	0.999
Linear		0.620	0.903	0.003	0.368	0.002	0.764
Quadratic		0.891	0.541	0.031	0.410	<0.001	0.900
Regression equation, R² value
Linear	Equation^e	y = 0.0035x + 9.4996	y = −0.001x + 9.1084	y = 0.0623x + 1.8484	y = −0.004x + 1.7347	y = −0.0168x + 2.6173	y = 0.0027x + 8.6649
Linear	R²	0.02	0.00	0.35	0.04	0.09	0.01
Quadratic	Equation^e	y = −0.0003x² + 0.0081x + 9.4873	y = −0.0018x² + 0.0244x + 9.0407	y = −0.0143x² + 0.2627x + 1.314	y = −0.0013x² + 0.0135x + 1.688	y = −0.0172x² + 0.2239x + 1.9757	y = 0.0004x²–0.0027x + 8.6793
Quadratic	R²	0.02	0.03	0.51	0.08	0.90	0.01

Notes: Data are expressed as mean (n = 3); Mean values in the same column with different superscripts differ significantly (p < 0.05).
Mean values in the same column with different superscripts differ significantly (p < 0.05).
Abbreviation: SEM, average standard error of means.
^a TCL40-TCL140, 40–140 g/kg dietary crude lipid.
^b Trypsin activity is expressed as units/min/mg protein at 37°C.
^c Amylase activity is expressed as micromole maltose released/min/mg protein at 37°C.
^d Lipase activity is expressed as units/min/mg protein at 37°C.
^e In equation, ‘x’ and ‘y’ represents dietary lipid levels and the respective parameters.

4 DISCUSSION

IGSW quality parameters of the present study like temperature, pH, DO, total alkalinity, total hardness, Ca²⁺, Mg²⁺ and K⁺ ions concentration were found within the recommended levels described by Talukdar et al. (2021) and Jahan et al. (2018) required for maximum growth performance of white-leg shrimp, P. vannamei in IGSW. The presence of higher levels of nitrogenous compounds in water like TA-N is highly toxic and lethal to shrimp (Burford & Lorenzen, 2004), causing severe stress, poor growth and even death of the cultured animal (Carbajal-Hernández et al., 2012). However, the level of toxicity also depends upon the size of the cultured animal, DO, temperature, pH, salinity and duration of exposure (Magallón Barajas et al., 2006). The level of TA-N was within the acceptable limit for P. vannamei culture (Talukdar et al., 2021). Nevertheless, TA-N is more lethal to shrimp than NO₃-N, and NO₂-N concentration in the surrounding water, whereas 100 mg/L of NO₂-N in water is reported to be deadly for shrimps (Van Rijn et al., 2006).

Previous studies on the dietary lipid requirement of P. vannamei explained that culture salinity and different growth stages could influence the dietary lipid requirement (Chen et al., 2014; González-Félix, Lawrence, et al., 2002; Hamidoghli et al., 2020; Jannathulla et al., 2019; Xie et al., 2019; Xu et al., 2018; Zhang et al., 2013; Zhu et al., 2010). In the current study, higher quadratic values of WG and SGR (R² = 0.97 and 0.98, respectively) were observed with the dietary lipid level up to 60 g/kg (TCL60) diet beyond this level growth reduced significantly. This agrees with the findings of González-Félix, Lawrence, et al. (2002) and González-Félix, Gatlin III, et al. (2002), who concluded that 30–90 g lipid/kg is optimum for higher survival, growth and production of juvenile P. vannamei at 25 ppt ambient salinity but growth performance gradually decreases at higher levels of dietary lipid. On the other hand, Hamidoghli et al. (2020) reported that P. vannamei juveniles require 56–60 g lipid/kg when reared in the biofloc system. The lowest FCR was recorded in the TCL60 group, but further enhancing the lipid levels beyond 60 g/kg significantly increased the value, suggesting efficient utilization of lipid using white-leg shrimp at this level to satisfy the energy requirement for higher growth, which is in agreement with Hamidoghli et al. (2020) and Jannathulla et al. (2019). In the current investigation, growth metrics and feed utilization using shrimp significantly reduced in higher lipid fed groups which might be attributed to increased metabolic stress on the cultured animal (Jannathulla et al., 2019). In contrast some authors (Xie et al., 2019; Xu et al., 2012; Zhang et al., 2013) recommended more than 100 g lipid/kg for optimum growth, survival, immune response and stress resistance of juvenile and post-larvae of whiteleg shrimp, P. vannamei under different culture salinities. In our present study, poor linear and quadratic values for survival and HPSI demonstrate that dietary lipids did not influence the survival rate and HPSI of P. vannamei juveniles reared in IGSW.

Contrast analysis of shrimp whole body composition indicated that feeding varying levels of dietary lipid has a significant effect on lipid and moisture contents in the body of shrimp. Shrimp whole-body lipid and moisture contents increased and decreased, respectively, with the dietary lipid level up to 140 g/kg. At the same time, shrimp whole-body ash and protein contents did not vary significantly among the treatment groups. Therefore, a diet with 400 g CP/kg and 60 g lipid/kg corresponding with 103.90 mg protein/Kcal DE P: E ratio could satisfy the protein and energy needs with higher body protein accretion at 15 ppt salinity. Moreover, at higher salinity regimes, energy requirement increases for osmoregulatory responses. Therefore energy satiation could be fulfilled using the protein-sparing action of non-protein nutrients to increase the somatic growth performance of shrimp. A similar observation was also made by Zhang et al. (2013), Xu et al. (2018) and Jannathulla et al. (2019) in P. vannamei juveniles. On the other hand, in contrast to our observation, Xie et al. (2019) and Hamidoghli et al. (2020) demonstrated significant variation in white shrimp's whole-body protein and ash contents due to feeding of graded levels of lipid.

Chen and Cheng (1995) and Adachi et al. (2003) described that subsequently after salinity adaptation, body amino acid pool derived amino acids are directly directed towards the synthesis of new tissues, osmoregulation, energy needs and immuno modulation. Pascual et al. (2003) also stated that the concentration of free amino acids in the hemolymph is very less as shrimps largely convert newly synthesized protein to hemocyanin (65%–95% of the hemolymph protein) after acclimatized to the ambient water salinity. In the present study, hemolymph hemocyanin and serum total protein contents did not differ significantly by feeding with different lipid levels. Depending upon the physiological status of shrimp, amino acids derived from lower dietary protein levels can be efficiently used either for osmoregulatory responses to metabolic energy needs. However, a diet with optimum levels of protein along with required levels of lipid could support the protein-sparing effect where dietary non-protein sources will supply the energy to meet the physiological energy demand, and a major portion of the amino acids will be directed towards the production and deposition of protein in the tissue of shrimp (Cuzon et al., 2004; Dall & Smith, 1986; Hamidoghli et al., 2018). Due to this reason, a diet with 400 g CP/kg with 60 g lipid/kg (TCL60) is attributed to maximum growth performance in shrimp. However, an excess dietary lipid and protein can cause a metabolic load on the shrimp resulting in growth retardation. This is in agreement with the observation by several authors (Hamidoghli et al., 2020; Xu et al., 2018) in P. vannamei.

Shan et al. (2019) described that a higher concentration of glucose in serum resulted in stress in shrimp associated with higher energy needs. In the present study, no significant difference was found in the serum glucose levels of shrimp while increasing the lipid levels of the diet. Shrimps have a restricted capability to convert glucose via glycolytic or glycogenesis pathways (Kumar et al., 2018; Rosas et al., 2000). Furthermore, shrimps could able to store very less quantity of carbohydrates in their body due to saturation of glycogenesis pathway at carbohydrate level 230 g/kg diet (Rosas et al., 2000). Concentrations of triglyceride and cholesterol in the serum are the key biomarkers of growth response in respect of diet design and formulations (Adhikari et al., 2004; Maheswaran et al., 2008). In the present study, total triglyceride and cholesterol contents in the serum increased significantly up to 60 g lipid/kg and then decreased significantly at higher lipid levels. Several authors well support this finding (Chen et al., 2014; Hamidoghli et al., 2020; Jannathulla et al., 2019; Xu et al., 2018) who described that dietary lipid at higher levels might likely reduce feed intake and poor feed utilization using shrimp P. vannamei resulting in lower total triglyceride and cholesterol contents in shrimp.

Among all the cultivable shrimp species, P. vannamei can easily adopt and withstand a wider ambient salinity ranges from 1 to 50 g/L, but optimum salinity is reported to be 15–30 ppt for better growth, survival rate and health status (Dall & Smith, 1981; Lightner et al., 2009; Pante, 1990). But deviation from the recommended culture salinity range results in breakdown of a significant amount of macro nutrients to meet the higher energy demand associated with the salinity induced osmoregulatory stress (Talukdar et al., 2021). These physiological adaptations lead to the reduced growth performance of shrimp and thus increase the aquaculture production cost. In the current study, no significant variation was observed in osmoregulatory capacity and osmolality of serum of the experimental shrimp. This may be because of hyper osmoregulation as a function of serum osmolality regulated through water salinity lesser than the iso-osmotic point (Castille Jr & Lawrence, 1981). In corroboration to our findings, Roy et al. (2007) stated that serum osmolality did not influence either dietary protein or lipid levels as it is a major function of water salinity. Thus, lower lipid levels in the diet may support the energy satiation for osmoregulatory adaptation as shrimp priorly ensures its physiological homeostasis due to deviation from the recommended optimum salinity range resulting in growth retardation of the shrimp.

In all the crustaceans, including shrimp, branchial Na⁺/K⁺-ATPase is directly involved in the osmoregulatory process and regulates the movement of ions throughout the body and the activation of the enzyme is directly related to the potassium ion concentration (Mantel & Farmer, 1983; Perry & Fryer, 1997). In our current study, no significant variation was found in Na⁺/K⁺-ATPase enzyme activities in the gill of the shrimp. This is attributed to the similar concentration of K⁺ ions in the rearing water after fortification with potash. In support of our findings, Hurtado et al. (2007) and Jana et al. (2021) also documented that dietary lipid or protein or the ambient water salinity did not influence the Na⁺/K⁺-ATPase enzyme activities in the gill of shrimp.

Proteolytic enzymes like trypsin and chymotrypsin play a major role in protein digestion as pepsin is absent in shrimps (Hernández & Murueta, 2009). However, Gaxiola et al. (2005) specified that requirement of dietary nutrients and availability of substrates in the digestive tract could alter the activation and function of digestive enzymes in shrimps. Hepatopancreatic amylase and lipase activities exhibited a significantly decreasing and increasing relationship, respectively, with the increasing dietary lipid levels which can be corroborated with the report of Lista and Velásquez (2003) who found that shrimps generally require lower levels of dietary lipid and higher levels beyond 100 g lipid/kg could not be sufficiently utilized. Similarly, Hamidoghli et al. (2020) concluded that feeding a lower dietary lipid level 45–90 g/kg diet could support the higher lipase activities in the hepatopancreas of shrimp and beyond that level the activity decreases gradually. Cuzon et al. (2004) and Li et al. (2008) also described the induced digestive enzyme activities for enhanced energy-dense nutrients to satiate the energy burden produced over iso-osmotic deviation stress. However, hepatopancreatic and intestinal trypsin activities did not differ significantly, which may be attributed to feeding similar dietary protein levels to different treatments.

5 CONCLUSION

From the current study, it can be recommended that a 55.80 to 58.80 g lipid/kg diet could be optimum for higher growth and production of white-leg shrimp, P. vannamei in 15 ppt IGSW. Feeding dietary lipid beyond this level reduced the growth performance. Nevertheless, the feed used for P. vannamei culture in the inland saline water is not yet standardized and the farmers are relying on regular commercial feeds available in the market. The outcome of the present study will serve as baseline information for the aqua feed industry to formulate an appropriately nutritious diet for sustainable shrimp culture in IGSW.

AUTHOR CONTRIBUTIONS

Prasanta Jana investigated the study, validated data, implemented software programs and prepared the original draft. Narottam Prasad Sahu conceptualized the study, supervised the study, and reviewed and edited the manuscript. Parimal Sardar designed methodology, involved in visualization and validation, and reviewed and edited the manuscript. Tincy Varghese and Ashutosh Dharmendra Deo performed data curation and validation. Nazeemashahul Shamna was involved in visualization and validation, and edited the manuscript. Vungurala Harikrishna supervised the study and edited the manuscript. Mritunjoy Paul performed investigation, formal analysis, software implementation and data curation. Nisha Chuphal performed formal analysis and data curation. Gopal Krishna reviewed and edited the manuscript.

ACKNOWLEDGEMENTS

The first author is grateful to the Indian Council of Agricultural Research for providing research grant for the PhD programme (2017-2020). The authors are thankful to Director of the Indian Council of Agricultural Research-Central Institute of Fisheries Education (ICAR-CIFE) for providing the necessary infrastructural facilities, and the World Bank, Govt. of India and ICAR funded National Agricultural Higher Education Project (NAHEP) are duly acknowledged for financial support to complete this research.

ETHICAL ISSUES

The authors hereby declare that there were no ethical issues while conducting this research work.

CONFLICT OF INTEREST

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Open Research

DATA AVAILABILITY STATEMENT

The data supporting this study's findings are available from the corresponding author upon reasonable request.

REFERENCES

Adachi, K., Hirata, T., Nishioka, T., & Sakaguchi, M. (2003). Hemocyte components in crustaceans convert hemocyanin into a phenoloxidase-like enzyme. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 134(1), 135–141. https://doi.org/10.1007/s00227-002-0995-2
10.1007/s00227?002?0995?2
PubMed Web of Science® Google Scholar
Adhikari, S., Sarkar, B., Chatterjee, A., Mahapatra, C. T., & Ayyappan, S. (2004). Effects of cypermethrin and carbofuran on certain hematological parameters and prediction of their recovery in a freshwater teleost, Labeo rohita (Hamilton). Ecotoxicology and Environmental Safety, 58(2), 220–226. https://doi.org/10.1016/j.ecoenv.2003.12.003
10.1016/j.ecoenv.2003.12.003
CAS PubMed Web of Science® Google Scholar
Aklakur, M. D. (2017). Nutritional intervention for sustainable production in inland saline aquaculture a budding perspective in India. Journal of Aquactic Marine Biology, 6, 172. https://doi.org/10.15406/jamb.2017.06.00172
10.15406/jamb.2017.06.00172
Google Scholar
Antony, J., Reddy, A. K., Sudhagar, A., Vungurala, H. K., & Roy, L. A. (2020). Effects of salinity on growth characteristics and osmoregulation of juvenile cobia, Rachycentron canadum (Linnaeus 1766), Reared in potassium-amended inland saline groundwater. Journal of the World Aquaculture Society, 52(1), 155–170. https://doi.org/10.1111/jwas.12741
10.1111/jwas.12741
Web of Science® Google Scholar
Antony, J., Vungurala, H., Saharan, N., Reddy, A. K., Chadha, N. K., Lakra, W. S., & Roy, L. A. (2015). Effects of salinity and Na⁺/K⁺ ratio on osmoregulation and growth performance of black tiger prawn, Penaeus monodon Fabricius, 1798, juveniles reared in inland saline water. Journal of the World Aquaculture Society, 46(2), 171–182. https://doi.org/10.1111/jwas.12179
10.1111/jwas.12179
CAS Web of Science® Google Scholar
AOAC (1995). In W. Horwitz (Ed.), Association of Official Analytical Chemistry (AOAC): Official methods of analysis of the Association of Official Analytical Chemists ( 15th ed., p. 1298). Association of Official Analytical Chemists.
Google Scholar
Arts, M. T., & Kohler, C. C. (2009). Health and condition in fish: The influence of lipids on membrane competency and immune response. In Lipids in aquatic ecosystems (pp. 237–256). Springer. https://doi.org/10.1007/978-0-387-89366-2_10
10.1007/978-0-387-89366-2_10
Web of Science® Google Scholar
Bou, M., Berge, G. M., Baeverfjord, G., Sigholt, T., Østbye, T. K., Romarheim, O. H., Hatlen, B., Leeuwis, R., Venegas, C., & Ruyter, B. (2017). Requirements of n-3 very long-chain PUFA in Atlantic salmon (Salmo salar L): Effects of different dietary levels of EPA and DHA on fish performance and tissue composition and integrity. British Journal of Nutrition, 117(1), 30–47. https://doi.org/10.1017/S0007114516004396
10.1017/S0007114516004396
CAS PubMed Web of Science® Google Scholar
Bui, H. T. T., Luu, T. Q., Fotedar, R., & Tantulo, U. (2017). Productivity of Sargassum linearifolium in potassium fortified inland saline water under laboratory conditions. Aquaculture Research, 48(11), 5631–5639. doi:10.1111/are.13385
10.1111/are.13385
CAS Web of Science® Google Scholar
Burford, M. A., & Lorenzen, K. (2004). Modeling nitrogen dynamics in intensive shrimp ponds: The role of sediment remineralization. Aquaculture, 229(1–4), 129–145. https://doi.org/10.1016/S0044-8486(03)00358-2
10.1016/S0044-8486(03)00358-2
CAS Web of Science® Google Scholar
Carbajal-Hernández, J. J., Sánchez-Fernández, L. P., Carrasco-Ochoa, J. A., & Martínez-Trinidad, J. F. (2012). Immediate water quality assessment in shrimp culture using fuzzy inference systems. Expert Systems with Applications, 39(12), 10571–10582. https://doi.org/10.1016/j.eswa.2012.02.141
10.1016/j.eswa.2012.02.141
Web of Science® Google Scholar
Castille, F. L., Jr., & Lawrence, A. L. (1981). The effect of salinity on the osmotic, sodium and chloride concentrations in the hemolymph of euryhaline shrimp of the genus Penaeus. Comparative Biochemistry and Physiology Part A: Physiology, 68(1), 75–80. https://doi.org/10.1016/0300-9629(81)90320-0
10.1016/0300?9629(81)90320?0
Web of Science® Google Scholar
Chen, J. C., & Cheng, S. Y. (1993a). Hemolymph PCO₂, hemocyanin, protein levels and urea excretions of Penaeus monodon exposed to ambient ammonia. Aquatic Toxicology, 27(3–4), 281–291. https://doi.org/10.1016/0166-445X(93)90059-A
10.1016/0166‐445X(93)90059‐A
CAS Web of Science® Google Scholar
Chen, J. C., & Cheng, S. Y. (1993b). Studies on haemocyanin and haemolymph protein levels of Penaeus japonicus based on sex, size and moulting cycle. Comparative Biochemistry and Physiology. B: Biochemistry and Molecular Biology, 106(2), 293–296.
10.1016/0305-0491(93)90303-M
Web of Science® Google Scholar
Chen, J. C., & Cheng, S. Y. (1995). Hemolymph oxygen content, oxyhemocyanin, protein levels and ammonia excretion in the shrimp Penaeus monodon exposed to ambient nitrite. Journal of Comparative Physiology B, 164(7), 530–535. https://doi.org/10.1007/BF00261393
10.1007/BF00261393
CAS Web of Science® Google Scholar
Chen, K., Li, E., Gan, L., Wang, X., Xu, C., Lin, H., Qin, J. G., & Chen, L. (2014). Growth and lipid metabolism of the pacific white shrimp Litopenaeus vannamei at different salinities. Journal of Shellfish Research, 33(3), 825–832. https://doi.org/10.2983/035.033.0317
10.2983/035.033.0317
Web of Science® Google Scholar
Cherry, I. S., & Crandall, L. A., Jr. (1932). The specificity of pancreatic lipase: Its appearance in the blood after pancreatic injury. American Journal of Physiology-Legacy Content, 100(2), 266–273. https://doi.org/10.1152/ajplegacy.1932.100.2.266
10.1152/ajplegacy.1932.100.2.266
CAS Google Scholar
Cuzon, G., Lawrence, A., Gaxiola, G., Rosas, C., & Guillaume, J. (2004). Nutrition of Litopenaeus vannamei reared in tanks or in ponds. Aquaculture, 235(1–4), 513–551. https://doi.org/10.1016/j.aquaculture.2003.12.022
10.1016/j.aquaculture.2003.12.022
CAS Web of Science® Google Scholar
Dall, W., & Smith, D. M. (1981). Ionic regulation of four species of penaeid prawn. Journal of Experimental Marine Biology and Ecology, 55(2–3), 219–232.
10.1016/0022-0981(81)90113-1
CAS Web of Science® Google Scholar
Dall, W., & Smith, D. M. (1986). Oxygen consumption and ammonia-N excretion in fed and starved tiger prawns, Penaeus esculentus Haswell. Aquaculture, 55(1), 23–33. https://doi.org/10.1016/0044-8486(86)90052-9
10.1016/0044-8486(86)90052-9
CAS Web of Science® Google Scholar
Davis, D. A., Boyd, C. E., Rouse, D. B., & Saoud, I. P. (2005). Effects of potassium, magnesium and age on growth and survival of Litopenaeus vannamei post-larvae reared in inland low salinity well waters in West Alabama. Journal of the World Aquaculture Society, 36(3), 416–419. https://doi.org/10.1111/j.1749-7345.2005.tb00346.x
10.1111/j.1749‐7345.2005.tb00346.x
Web of Science® Google Scholar
Diaz, F., Farfan, C., Sierra, E., & Re, A. D. (2001). Effects of temperature and salinity fluctuation on the ammonium excretion and osmoregulation of juveniles of Penaeus vannamei, Boone. Marine and Freshwater Behaviour and Physiology, 34(2), 93–104. https://doi.org/10.1080/10236240109379062
10.1080/10236240109379062
CAS Web of Science® Google Scholar
FAO. (2018). The state of the world fisheries and aquaculture. Food and Agricultural Organization of the United Nations.
Google Scholar
FAO. (2020). The state of world fisheries and aquaculture. Sustainability in action. Food and Agriculture Organization of the United Nations.
Google Scholar
Flores, M., Díaz, F., Medina, R., Re, A. D., & Licea, A. (2007). Physiological, metabolic and haematological responses in white shrimp Litopenaeus vannamei (Boone) juveniles fed diets supplemented with astaxanthin acclimated to low-salinity water. Aquaculture Research, 38(7), 740–747. https://doi.org/10.1111/j.1365-2109.2007.01720.x
10.1111/j.1365‐2109.2007.01720.x
CAS Web of Science® Google Scholar
Garcia-Carreno, F. L., Hernandez-Cortes, M. P., & Haard, N. F. (1994). Enzymes with peptidase and proteinase activity from the digestive systems of a freshwater and a marine decapod. Journal of Agricultural and Food Chemistry, 42(7), 1456–1461. https://doi.org/10.1021/jf00043a013
10.1021/jf00043a013
CAS Web of Science® Google Scholar
Gaxiola, G., Cuzon, G., García, T., Taboada, G., Brito, R., Chimal, M. E., Paredes, A., Soto, L., Rosas, C., & Van Wormhoudt, A. (2005). Factorial effects of salinity, dietary carbohydrate and moult cycle on digestive carbohydrases and hexokinases in Litopenaeus vannamei (Boone, 1931). Comparative Biochemistry and Physiology Part A: Molecular and Integrative Physiology, 140(1), 29–39. https://doi.org/10.1016/j.cbpb.2004.10.018
10.1016/j.cbpb.2004.10.018
PubMed Web of Science® Google Scholar
Goda, A. M. S. (2008). Effect of dietary protein and lipid levels and protein–energy ratio on growth indices, feed utilization and body composition of freshwater prawn, Macrobrachium rosenbergii (de man 1879) post larvae. Aquaculture Research, 39(8), 891–901. https://doi.org/10.1111/j.1365-2109.2008.01947.x
10.1111/j.1365‐2109.2008.01947.x
CAS Web of Science® Google Scholar
González-Félix, M. L., da Silva, F. S. D., Davis, D. A., Samocha, T. M., Morris, T. C., Wilkenfeld, J. S., & Perez-Velazquez, M. (2010). Replacement of fish oil in plant based diets for Pacific white shrimp (Litopenaeus vannamei). Aquaculture, 309(1–4), 152–158. https://doi.org/10.1016/j.aquaculture.2010.08.028
10.1016/j.aquaculture.2010.08.028
CAS Web of Science® Google Scholar
González-Félix, M. L., Gatlin, D. M., III, Lawrence, A. L., & Perez-Velazquez, M. (2002). Effect of dietary phospholipid on essential fatty acid requirements and tissue lipid composition of Litopenaeus vannamei juveniles. Aquaculture, 207(1–2), 151–167. https://doi.org/10.1016/S0044-8486(01)00797-9
10.1016/S0044-8486(01)00797-9
CAS Web of Science® Google Scholar
González-Félix, M. L., Lawrence, A. L., Gatlin, D. M., III, & Perez-Velazquez, M. (2002). Growth, survival and fatty acid composition of juvenile Litopenaeus vannamei fed different oils in the presence and absence of phospholipids. Aquaculture, 205(3–4), 325–343. https://doi.org/10.1016/S0044-8486(01)00684-6
10.1016/S0044?8486(01)00684?6
CAS Web of Science® Google Scholar
Hamidoghli, A., Won, S., Aya, F. A., Yun, H., Bae, J., Jang, I. K., & Bai, S. C. (2020). Dietary lipid requirement of whiteleg shrimp Litopenaeus vannamei juveniles cultured in biofloc system. Aquaculture Nutrition, 26(3), 603–612. https://doi.org/10.1111/anu.13021
10.1111/anu.13021
CAS Web of Science® Google Scholar
Hamidoghli, A., Yun, H., Shahkar, E., Won, S., Hong, J., & Bai, S. C. (2018). Optimum dietary protein-to-energy ratio for juvenile whiteleg shrimp, Litopenaeus vannamei, reared in a biofloc system. Aquaculture Research, 49(5), 1875–1886. https://doi.org/10.1111/are.13643
10.1111/are.13643
CAS Web of Science® Google Scholar
He, H., Lawrence, A. L., & Liu, R. (1992). Evaluation of dietary essentiality of fat-soluble vitamins, a, D, E and K for penaeid shrimp (Penaeus vannamei). Aquaculture, 103(2), 177–185. https://doi.org/10.1016/0044-8486(92)90411-D
10.1016/0044-8486(92)90411-D
Web of Science® Google Scholar
Hernández, J. C. S., & Murueta, J. H. C. (2009). Activity of trypsin from Litopenaeus vannamei. Aquaculture, 290(3–4), 190–195. https://doi.org/10.1016/j.aquaculture.2009.02.034
10.1016/j.aquaculture.2009.02.034
Web of Science® Google Scholar
Huong, D. T. T., Jasmani, S., Jayasankar, V., & Wilder, M. (2010). Na/KATPase activity and osmo-ionic regulation in adult whiteleg shrimp Litopenaeus vannamei exposed to low salinities. Aquaculture, 304, 88–94. https://doi.org/10.1016/j.aquaculture.2010.03.025
10.1016/j.aquaculture.2010.03.025
CAS Web of Science® Google Scholar
Hurtado, M. A., Racotta, I. S., Civera, R., Ibarra, L., Hernández-Rodríguez, M., & Palacios, E. (2007). Effect of hypo-and hypersaline conditions on osmolality and Na⁺/K⁺-ATPase activity in juvenile shrimp (Litopenaeus vannamei) fed low-and high-HUFA diets. Comparative Biochemistry and Physiology Part A: Molecular and Integrative Physiology, 147(3), 703–710. https://doi.org/10.1016/j.cbpa.2006.07.002
10.1016/j.cbpa.2006.07.002
CAS PubMed Web of Science® Google Scholar
Jahan, I., Reddy, A. K., Sudhagar, S. A., Harikrishna, V., Singh, S., Varghese, T., & Srivastava, P. P. (2018). The effect of fortification of potassium and magnesium in the diet and culture water on growth, survival and osmoregulation of Pacific white shrimp, Litopenaeus vannamei reared in inland ground saline water. Turkish Journal of Fisheries and Aquatic Sciences, 18(10), 1235–1243. https://doi.org/10.4194/1303-2712-v18_10_10
10.4194/1303‐2712‐v18_10_10
Web of Science® Google Scholar
Jamil, A., Riaz, S., Ashraf, M., & Foolad, M. R. (2011). Gene expression profiling of plants under salt stress. Critical Reviews in Plant Sciences, 30(5), 435–458. https://doi.org/10.1080/07352689.2011.605739
10.1080/07352689.2011.605739
Web of Science® Google Scholar
Jana, P., Prasad Sahu, N., Sardar, P., Shamna, N., Varghese, T., Dharmendra Deo, A., Harikrishna, V., Paul, M., Panmei, H., Gupta, G., Nanda, C., & Krishna, G. (2021). Dietary protein requirement of white shrimp, Penaeus vannamei (Boone, 1931) juveniles, reared in inland ground water of medium salinity. Aquaculture Research, 52(6), 2501–2517. https://doi.org/10.1111/are.15100
10.1111/are.15100
CAS Web of Science® Google Scholar
Jannathulla, R., Chitra, V., Vasanthakumar, D., Nagavel, A., Ambasankar, K., Muralidhar, M., & Dayal, J. S. (2019). Effect of dietary lipid/essential fatty acid level on Pacific whiteleg shrimp, Litopenaeus vannamei (Boone, 1931) reared at three different water salinities–emphasis on growth, hemolymph indices and body composition. Aquaculture, 513, 734405. https://doi.org/10.1016/j.aquaculture.2019.734405
10.1016/j.aquaculture.2019.734405
CAS Web of Science® Google Scholar
Kumar, V., Habte-Tsion, H. M., Allen, K. M., Bowman, B. A., Thompson, K. R., El-Haroun, E., Filer, K., & Tidwell, J. H. (2018). Replacement of fish oil with Schizochytrium meal and its impacts on the growth and lipid metabolism of Pacific white shrimp (Litopenaeus vannamei). Aquaculture Nutrition, 24(6), 1769–1781. https://doi.org/10.1111/anu.12816
10.1111/anu.12816
CAS Web of Science® Google Scholar
Li, E. C., Chen, L. Q., Zeng, C., Xion, Z. Q., Lin, C., Sun, X. J., & Sen, H. L. (2008). Effects of dietary protein levels on growth, survival, body composition and hepatopancreas histological structure of the white shrimp, Litopenaeus vannamei, at different ambient salinities. Journal of Fisheries of China, 32, 425–434.
Google Scholar
Li, E., Wang, X., Chen, K., Xu, C., Qin, J. G., & Chen, L. (2017). Physiological change and nutritional requirement of Pacific white shrimp Litopenaeus vannamei at low salinity. Reviews in Aquaculture, 9(1), 57–75. https://doi.org/10.1111/raq.12104
10.1111/raq.12104
Web of Science® Google Scholar
Lightner, D. V., Redman, R. M., Arce, S. M., & Moss, M. S. (2009). Specific pathogen free shrimp stocks in shrimp farming facilities as a novel method for disease control in crustaceans. In S. E. Shumway & G. E. Rodrick (Eds.), Shellfish safety and quality (pp. 384–424). CRC Press, Woodhead Publishing Limited. https://doi.org/10.1533/9781845695576.3.384
10.1533/9781845695576.3.384
Google Scholar
Lignot, J. H., Spanings-Pierrot, C., & Charmantier, G. (2000). Osmoregulatory capacity as a tool in monitoring the physiological condition and the effect of stress in crustaceans. Aquaculture, 191(1–3), 209–245. https://doi.org/10.1016/S0044-8486(00)00429-4
10.1016/S0044-8486(00)00429-4
CAS Web of Science® Google Scholar
Lista, M., & Velásquez, C. (2003). Influence of three experimental diets in the growth of postlarvae of Litopenaeus vannamei (Boone, 1931). Revista Científica, 13, 167–172.
Web of Science® Google Scholar
Lowry, O. H., Rosebrough, N. J., Farr, A. L., & Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry, 193, 265–275.
10.1016/S0021-9258(19)52451-6
CAS PubMed Web of Science® Google Scholar
Luo, Z. H. I., Liu, Y. J., Mai, K. S., Tian, L. X., Liu, D. H., Tan, X. Y., & Lin, H. Z. (2005). Effect of dietary lipid level on growth performance, feed utilization and body composition of grouper Epinephelus coioides juveniles fed isonitrogenous diets in floating netcages. Aquaculture International, 13(3), 257–269. https://doi.org/10.1007/s10499-004-2478-6
10.1007/s10499?004?2478?6
CAS Web of Science® Google Scholar
Magallón Barajas, F., Villegas, R. S., Clark, G. P., Mosqueda, J. G., & Moreno, B. L. (2006). Daily variation in short-term static toxicity of unionized ammonia in Litopenaeus vannamei (Boone) postlarvae. Aquaculture Research, 37(14), 1406–1412. https://doi.org/10.1111/j.1365-2109.2006.01573.x
10.1111/j.1365‐2109.2006.01573.x
CAS Web of Science® Google Scholar
Maheswaran, R., Devapaul, A., Muralidharan, S., Velmurugan, B., & Ignacimuthu, S. (2008). Haematological studies of freshwater fish, Clarias batrachus (L.) exposed to mercuric chloride. International Journal of Integrated Biology, 2, 49–54.
CAS Google Scholar
Mantel, L. H., & Farmer, L. L. (1983). Osmotic and ionic regulation. In D. E. Bliss & L. H. Mantel (Eds.), The biology of crustacea (Vol. 5, pp. 54–126). Academic Press.
Google Scholar
McCormick, S. D. (1993). Methods for nonlethal gill biopsy and measurement of Na+K+-ATPase activity. Canadian Journal of Fisheries and Aquatic Sciences, 50(3), 656–658. https://doi.org/10.1139/f93-075
10.1139/f93?075
CAS Web of Science® Google Scholar
Pante, M. J. R. (1990). Influence of environmental stress on the heritability of molting frequency and growth rate of the penaeid shrimp, Penaeus vannamei. University of Houston-Clear lake, Houston, TX, USA, M.Sc. Thesis.
Google Scholar
Partridge, G. J., Lymbery, A. J., & Bourke, D. K. (2008). Larval rearing of barramundi (Lates calcarifer) in saline groundwater. Aquaculture, 278(1–4), 171–174. https://doi.org/10.1016/j.aquaculture.2008.03.023
10.1016/j.aquaculture.2008.03.023
CAS Web of Science® Google Scholar
Pascual, C., Gaxiola, G., & Rosas, C. (2003). Blood metabolites and hemocyanin of the white shrimp, Litopenaeus vannamei: The effect of culture conditions and a comparison with other crustacean species. Marine Biology, 142(4), 735–745. https://doi.org/10.1007/s00227-002-0995-2
10.1007/s00227‐002‐0995‐2
CAS Web of Science® Google Scholar
Perry, S. F., & Fryer, J. N. (1997). Proton pumps in the fish gill and kidney. Fish Physiology and Biochemistry, 17(1–6), 363–369. https://doi.org/10.1023/A:1007746217349
10.1023/A:1007746217349
CAS Web of Science® Google Scholar
Raizada, S., Purushothaman, C. S., Sharma, V. K., Harikrishna, V., Rahaman, M., Agrahari, R. K., Venugopal, G., & Kumar, A. (2015). Survival and growth of tiger shrimp (Penaeus monodon) in inland saline water supplemented with potassium. Proceedings of the National Academy of Sciences, India Section B: Biological Sciences, 85(2), 491–497. https://doi.org/10.1007/s40011-014-0372-1
10.1007/s40011‐014‐0372‐1
CAS Google Scholar
Rick, W., & Stegbauer, H. P. (1974). Amylase measurement of reducing groups. In H. V. Bergmeyer (Ed.), Methods of enzymatic analysis (pp. 885–889). Academic Press. https://doi.org/10.1016/B978-0-12-091302-2.50074-8
10.1016/B978-0-12-091302-2.50074-8
Google Scholar
Rosas, C., Cuzon, G., Gaxiola, G., Arena, L., Lemaire, P., Soyez, C., & Van Wormhoudt, A. (2000). Influence of dietary carbohydrate on the metabolism of juvenile Litopenaeus stylirostris. Journal of Experimental Marine Biology and Ecology, 249(2), 181–198. https://doi.org/10.1016/S0022-0981(00)00184-2
10.1016/S0022-0981(00)00184-2
CAS PubMed Web of Science® Google Scholar
Roy, L. A., Davis, D. A., Saoud, I. P., & Henry, R. P. (2007). Effects of varying levels of aqueous potassium and magnesium on survival, growth, and respiration of the Pacific white shrimp, Litopenaeus vannamei, reared in low salinity waters. Aquaculture, 262(2–4), 461–469. https://doi.org/10.1016/j.aquaculture.2006.10.011
10.1016/j.aquaculture.2006.10.011
CAS Web of Science® Google Scholar
Samocha, T. M., Hamper, L., Emberson, C. R., Davis, A. D., McIntosh, D., Lawrence, A. L., & Van Wyk, P. M. (2002). Review of some recent developments in sustainable shrimp farming practices in Texas, Arizona, and Florida. Journal of Applied Aquaculture, 12(1), 1–42. https://doi.org/10.1300/J028v12n01_01
10.1300/J028v12n01_01
Google Scholar
Saoud, I. P., Davis, D. A., & Rouse, D. B. (2003). Suitability studies of inland well waters for Litopenaeus vannamei culture. Aquaculture, 217(1–4), 373–383. https://doi.org/10.1016/S0044-8486(02)00418-0
10.1016/S0044‐8486(02)00418‐0
Web of Science® Google Scholar
Saoud, I. P., Roy, L. A., & Davis, D. A. (2007). Chelated potassium and arginine supplementation in diets of Pacific white shrimp reared in low-salinity waters of West Alabama. North American Journal of Aquaculture, 69(3), 265–270. https://doi.org/10.1577/A06-045.1
10.1577/A06‐045.1
Web of Science® Google Scholar
Saraswathy, R., Muralidhar, M., Balasubramanian, C. P., Rajesh, R., Sukumaran, S., Kumararaja, P., & Vijayan, K. K. (2021). Osmo-ionicregulation in whiteleg shrimp, Penaeus vannamei, exposed to climate change-induced low salinities. Aquaculture Research, 52(2), 771–782. https://doi.org/10.1111/are.14933
10.1111/are.14933
CAS Web of Science® Google Scholar
Shan, H., Wang, T., Dong, Y., & Ma, S. (2019). Effects of dietary Ampithoe sp. supplementation on the growth, energy status, antioxidant capacity, and ammonia-N tolerance of the shrimp Litopenaeus vannamei: Continuous versus interval feeding. Aquaculture, 509, 32–39. https://doi.org/10.1016/j.aquaculture.2019.05.021
10.1016/j.aquaculture.2019.05.021
CAS Web of Science® Google Scholar
Singha, K. P., Shamna, N., Sahu, N. P., Sardar, P., HariKrishna, V., Thirunavukkarasar, R., Kumar, M., & Krishna, G. (2020). Feeding graded levels of protein to genetically improved farmed tilapia (GIFT) juveniles reared in inland saline water: Effects on growth and gene expression of IGFI, IGF-IR and IGF-BPI. Aquaculture, 735306, 735306. https://doi.org/10.1016/j.aquaculture.2020.735306
10.1016/j.aquaculture.2020.735306
Web of Science® Google Scholar
Talukdar, A., Deo, A. D., Sahu, N. P., Sardar, P., Aklakur, M., Prakash, S., Shamna, N., & Kumar, S. (2020). Effects of dietary protein on growth performance, nutrient utilization, digestive enzymes and physiological status of grey mullet, Mugil cephalus L. fingerlings reared in inland saline water. Aquaculture Nutrition, 26(3), 921–935. https://doi.org/10.1111/anu.13050
10.1111/anu.13050
CAS Web of Science® Google Scholar
Talukdar, A., Dharmendra Deo, A., Prasad Sahu, N., Sardar, P., Aklakur, M., Harikrishna, V., Prakash, S., Shamna, N., & Jana, P. (2021). Effects of different levels of dietary protein on the growth performance, nutrient utilization, digestive enzymes and physiological status of white shrimp, Litopenaeus vannamei juveniles reared in inland saline water. Aquaculture Nutrition, 27(1), 77–90. https://doi.org/10.1111/anu.13166
10.1111/anu.13166
CAS Web of Science® Google Scholar
Tantulo, U., & Fotedar, R. (2006). Comparison of growth, osmoregulatory capacity, ionic regulation and organosomatic indices of black tiger prawn (Penaeus monodon Fabricius, 1798) juveniles reared in potassium fortified inland saline water and ocean water at different salinities. Aquaculture, 258(1–4), 594–605. https://doi.org/10.1016/j.aquaculture.2006.04.038
10.1016/j.aquaculture.2006.04.038
CAS Web of Science® Google Scholar
Toledo, T. M., Silva, B. C., Vieira, F. D. N., Mouriño, J. L. P., & Seiffert, W. Q. (2016). Effects of different dietary lipid levels and fatty acids profile in the culture of white shrimp Litopenaeus vannamei (Boone) in biofloc technology: Water quality, biofloc composition, growth and health. Aquaculture Research, 47(6), 1841–1851. https://doi.org/10.1111/are.12642
10.1111/are.12642
CAS Web of Science® Google Scholar
Tseng, Y. C., & Hwang, P. P. (2008). Some insights into energy metabolism for osmoregulation in fish. Comparative Biochemistry and Physiology Part C: Toxicology and Pharmacology, 148(4), 419–429. https://doi.org/10.1016/j.cbpc.2008.04.009
10.1016/j.cbpc.2008.04.009
PubMed Web of Science® Google Scholar
Tzeng, B. S., Lin, M. N., & Sheen, S. S. (2004). Effect of dietary lipid levels on the reproductive performance of white shrimp (Litopenaeus vannamei). Journal of Taiwan Fisheries Research, 12, 43–48.
Google Scholar
Ur-Rahman, S., Jain, A. K., Reddy, A. K., Kumar, G., & Raju, K. D. (2005). Ionic manipulation of inland saline groundwater for enhancing survival and growth of Penaeus monodon (Fabricius). Aquaculture Research, 36(12), 1149–1156. https://doi.org/10.1111/j.1365-2109.2005.01322.x
10.1111/j.1365‐2109.2005.01322.x
Web of Science® Google Scholar
Van Rijn, J., Tal, Y., & Schreier, H. J. (2006). Denitrification in recirculating systems: Theory and applications. Aquacultural Engineering, 34(3), 364–376. https://doi.org/10.1016/j.aquaeng.2005.04.004
10.1016/j.aquaeng.2005.04.004
Web of Science® Google Scholar
Vargas-Albores, F., Guzmán, M. A., & Ochoa, J. L. (1993). An anticoagulant solution for haemolymph collection and prophenoloxidase studies of penaeid shrimp (Penaeus californiensis). Comparative Biochemistry and Physiology Part A: Physiology, 106(2), 299–303. https://doi.org/10.1016/0300-9629(93)90516-7
10.1016/0300?9629(93)90516?7
Web of Science® Google Scholar
Wang, X. D., Li, E. C., Wang, S. F., Qin, J. G., Chen, X. F., Lai, Q. M., Chen, C., Xu, L., Gan, N., Yu, Z. Y., & Du, Z. Y. (2015). Protein-sparing effect of carbohydrate in the diet of white shrimp Litopenaeus vannamei at low salinity. Aquaculture Nutrition, 21(6), 904–912. https://doi.org/10.1111/anu.12221
10.1111/anu.12221
CAS Web of Science® Google Scholar
Wang, X., Li, E., Qin, J. G., Wang, S., Chen, X., Cai, Y., Chen, K., Gan, L., Yu, N., Du, Z.-Y., & Du, Z. (2014). Growth, body composition, and ammonia tolerance of juvenile white shrimp Litopenaeus vannamei fed diets containing different carbohydrate levels at low salinity. Journal of Shellfish Research, 33(2), 511–517. https://doi.org/10.2983/035.033.0220
10.2983/035.033.0220
CAS Web of Science® Google Scholar
Xie, S., Wei, D., Fang, W., Wan, M., Guo, T., Liu, Y., Yin, P., Tian, L., & Niu, J. (2019). Optimal dietary lipid requirement of postlarval white shrimp, Litopenaeus vannamei in relation to growth performance, stress tolerance and immune response. Aquaculture Nutrition, 25(6), 1231–1240. https://doi.org/10.1111/anu.12937
10.1111/anu.12937
CAS Web of Science® Google Scholar
Xu, C., Li, E., Liu, Y., Wang, S., Wang, X., Chen, K., Qin, L., & Chen, L. (2018). Effect of dietary lipid level on growth, lipid metabolism and health status of the Pacific white shrimp Litopenaeus vannamei at two salinities. Aquaculture Nutrition, 24(1), 204–214. https://doi.org/10.1111/anu.12548
10.1111/anu.12548
CAS Web of Science® Google Scholar
Xu, W. J., Morris, T. C., & Samocha, T. M. (2016). Effects of C/N ratio on biofloc development, water quality, and performance of Litopenaeus vannamei juveniles in a biofloc-based, high-density, zero-exchange, outdoor tank system. Aquaculture, 453, 169–175. https://doi.org/10.1016/j.aquaculture.2015.11.021
10.1016/j.aquaculture.2015.11.021
CAS Web of Science® Google Scholar
Xu, W. J., Pan, L. Q., Zhao, D. H., & Huang, J. (2012). Preliminary investigation into the contribution of bioflocs on protein nutrition of Litopenaeus vannamei fed with different dietary protein levels in zero-water exchange culture tanks. Aquaculture, 350, 147–153. https://doi.org/10.1016/j.aquaculture.2012.04.003
10.1016/j.aquaculture.2012.04.003
Web of Science® Google Scholar
Yun, H., Shahkar, E., Katya, K., Jang, I. K., Kim, S. K., & Bai, S. C. (2016). Effects of bioflocs on dietary protein requirement in juvenile whiteleg shrimp, Litopenaeus vannamei. Aquaculture Research, 47(10), 3203–3214. https://doi.org/10.1111/are.12772
10.1111/are.12772
CAS Web of Science® Google Scholar
Zhang, S. P., Li, J. F., Wu, X. C., Zhong, W. J., Xian, J. A., Liao, S. A., Miao, Y. T., & Wang, A. L. (2013). Effects of different dietary lipid level on the growth, survival and immune-relating genes expression in Pacific white shrimp, Litopenaeus vannamei. Fish and Shellfish Immunology, 34(5), 1131–1138. https://doi.org/10.1016/j.fsi.2013.01.016
10.1016/j.fsi.2013.01.016
CAS PubMed Web of Science® Google Scholar
Zhou, X. X., Zhang, J. Y., Liu, S. L., & Ding, Y. T. (2014). Supplementation of sodium chloride in diets to improve the meat quality of Pacific white shrimp, Litopenaeus vannamei, reared in low-salinity water. Aquaculture Research, 45(7), 1187–1195. https://doi.org/10.1111/are.12062
10.1111/are.12062
CAS Web of Science® Google Scholar
Zhu, X. Z., Liu, Y. J., Tian, L. X., Mai, K. S., Zheng, S. X., Pan, Q. J., Cai, M., Zheng, C., Zhang, Q., & Hu, Y. (2010). Effects of dietary protein and lipid levels on growth and energy productive value of pacific white shrimp, Litopenaeus vannamei, at different salinities. Aquaculture Nutrition, 16(4), 392–399. https://doi.org/10.1111/j.1365-2095.2009.00677.x
10.1111/j.1365?2095.2009.00677.x
CAS Web of Science® Google Scholar

Citing Literature

All articles

Dietary lipid requirement of juvenile white-leg shrimp, Penaeus vannamei (Boone, 1931) reared in inland ground saline water of 15 ppt

Abstract

1 INTRODUCTION

2 MATERIALS AND METHODS

2.1 Ethical declarations

2.2 Lab animal

2.3 Formulation and preparation of diets

2.4 Collection, filling and storage of IGSW

2.5 Setting up of experiment and feeding trial

2.6 Sampling

2.7 Proximate composition of the diets and tissues

2.8 Growth metrics, nutrient utilization and survival

2.9 Hemato-biochemical analysis

2.10 Enzyme assay

2.10.1 Quantification of tissue protein

2.10.2 Activities of Na+/K+-ATPase

2.10.3 Activities of digestive enzymes

2.11 Statistical analysis

3 RESULTS

3.1 Growth metrics, nutrient utilization and survivability of shrimp

3.2 Proximate composition of the whole body of shrimp

3.3 Hemolymph hemocyanin and serum biochemical indices

3.4 Na+/K+-ATPase enzyme activity, osmolality and osmoregulatory capacity (OC) of shrimp

3.5 Hepatopancreatic and intestinal digestive enzymes activities

4 DISCUSSION

5 CONCLUSION

AUTHOR CONTRIBUTIONS

ACKNOWLEDGEMENTS

ETHICAL ISSUES

CONFLICT OF INTEREST

Open Research

DATA AVAILABILITY STATEMENT

REFERENCES

Citing Literature

Figures

References

Related

Information

2.10.2 Activities of Na⁺/K⁺-ATPase

3.4 Na⁺/K⁺-ATPase enzyme activity, osmolality and osmoregulatory capacity (OC) of shrimp