Performance of Dried Distillery Grain Waste on the Growth and Survival of Rohu (Labeo rohita) Larvae

Padala Dharmakar1*, S. Aanand1, J. Stephen Sampath Kumar2,

Muralidhar P. Ande3, P. Padmavathy4, J. Jaculine Pereira4 and Ch. Balakrishna5

1Erode Bhavanisagar Centre for Aquaculture, Tamil Nadu Dr. J. Jayalalithaa Fisheries University, Bhavanisagar, India

2Directorate of Sustainable Aquaculture, Tamil Nadu Dr. J. Jayalalithaa Fisheries University, Nagapattinam, India.

3ICAR-Central Institute of Fisheries Education, Kakinada Center, Kakinada, India

4Fisheries College and Research Institute, Tamil Nadu Dr. J. Jayalalithaa Fisheries University, Thoothukudi, India.

5ICAR-Krishi Vigyan Kendra, Acharya N G Ranga Agricultural University, Amadalavalasa, India.

ABSTRACT

Fish meals followed by oil cakes are the primary protein sources in any aquafeed formulation. The higher-cost of oil cakes and uncertainty in the fish meal (FM) availability led to the search for good quality alternative feed ingredients. By-product protein sources are cheaper and readily available to substitute the expensive ingredients in traditional feed production. The present study was carried out to investigate the performance of dried distiller’s grain waste (DDGW) by partial replacement of groundnut oil cake (GNOC) on the growth and survival of Labeo rohita in the nursery phase. Three isonitrogenous diets incorporating DDGW at (T1) 30%, (T2) 40% and (T3) 50% kg/feed replacement of GNOC were used as test diets with and diets only GNOC was taken as the control (C). The experimental trial was conducted in duplicate groups of 3000 rohu juvenile (mean initial weight 0.04±0.03 g) and was fed with the diets for 60 days. The growth performance and feed utilization parameters, viz., weight gain % (WG), specific growth rate (SGR), protein efficiency ratio (PER) and feed conversion ratio (FCR), were recorded. The fishes’ whole body proximate composition and amino acid profiles were not significantly different among experimental diets. The results revealed that dietary DDGW @50% replacement of GNOC had shown the higher final weight and specific growth rate. Feed conversion and protein efficiency rates were similar among the dietary treatments. The digestive enzyme activity remained unaffected except for amylase activity, which increased significantly in the 50% DDGW replacement group. The present study indicates that DDGW can replace GNOC without affecting feed utilization parameters for better growth performance and digestive enzyme activity in the diet of L. rohita.

Article Information

Received 14 May 2023

Revised 05 August 2023

Accepted 24 August 2023

Available online 11 November 2023

(early access)

Published 29 March 2025

Authors’ Contribution

This study was conducted in cooperation between all authors. PD: Investigation and conducted the feeding trial, Analyzed the samples for proximate and amino acid, writing drafted the original manuscript, SA: Designed the study, Analysed the results, and reviewed the manuscript. Supervision, JSSK: Investigation, Supervision, APM: Technical and Statistical support, and reviewed the manuscript, PP: reviewed the manuscript, JJP: reviewed the manuscript and analysed the results and CHB: Data analysis, manuscript review and editing.

Key words

Dried distiller’s grains waste, Growth performance, Labeo rohita, Digestive enzyme

DOI: https://dx.doi.org/10.17582/journal.pjz/20230514060530

* Corresponding author: dharmabfsc@gmail.com

0030-9923/2025/0002-0777 $ 9.00/00

Copyright 2025 by the authors. Licensee Zoological Society of Pakistan.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

INTRODUCTION

The rapid expansion of aquaculture among food-producing sectors was due to the demand created the increasing population growth and over-exploitation of existing resources. The farm production of fish has continuously grown to sustain the demand gap (Lahsen and Iddya, 2014). The global average per capita fish consumption rose from 17.4 to 20.5 kg annually (FAO, 2022). Further expansion of the aquaculture industry was attributed to advanced technology, improved farming methods, and high-quality fish feed production (Tran and Rodela, 2019). Carp culture forms the backbone of freshwater aquaculture practice in India, contributing to about 87% of total production. The improved feed management practice in the culture of indian major carps can successfully fill the gap between productivity and potential. The nutrient and feed inputs supply will grow equally to maintain its current growth rate. In contrast, feed ingredient availability for aquaculture remains static, and other sectors compete for the same feed resources (Tacon and Metian, 2015).

Carp culture largely depends on the quality of feeds, but ensuring the availability of these feed ingredients remains a challenge for the Indian aquaculture industry. The key aquafeed constituents like fish meal, soy meal and oilseed cakes are in a race with native animal husbandry (Hasan, 2007). Plant-based oilseed meals processed in traditional methods cannot be utilized at high levels (Soltan et al., 2015). Contaminants like mycotoxins may also limit vegetable feedstuffs in aquafeeds (Hendricks, 2003). Among the oilcake diets, groundnut oil cake (GNOC) is widely used feed ingredient in nursery rearing of carps, but deficient in methionine, tryptophan and tyrosine (Singh et al., 1981). Its storage duration and quality are poor as it may develop aflatoxins in the long run (Girma et al., 2011). Therefore, partially replacing of these ingredients with alternate feed ingredients like millets, distiller’s dried grains waste (DDGW) and other brewery waste ingredients containing low phytate and highly digestible nutrient composition seems promising for aquaculture feed industry. However, the potential of these alternate feed ingredients needs to be explored based on their price, availability, and nutritive value (Ravindran, 2013). The basic knowledge of small-scale farmers on fish feed needs to be enlightened to enhance aquaculture production. In practice, the concept of complete feed has not reached the fish culturist, unlike shrimp farmers. So to find an alternative source of protein with some better characteristics, cost or performance are considered for an expected success. One solution to meeting the protein requirement of cultured species is the utilization of by-products from the brewing industry, which are natural diet additives that have shown better growth on a few fish species (Rumsey et al., 1991; Oliva-Teles and Goncalves, 2001).

The brewing industry tends to be more environmentally friendly (Ishiwaki et al., 2000). The dried distiller’s grain waste (DDGW) is a by-product of the spirit/brewery industry during bio-ethanol production. Spent grain is the most abundant brewery by-product, contributing to 85% of the total by-products generated (Reinold, 1997). The continuous large-scale production by industries has led to a significant increase in waste generation, causing detrimental environmental pollution. Utilization of these wastes as animal feed is an economical alternative. Brewers spent grain is a high-value product containing hemicelluloses, lignin, proteins, and sugars (Buffington, 2014).

Earlier studies among a few fishes showed that brewery waste-activated sludge (BWAS) could be added to trout diets up to 10% (Windell, 1974). Bays (1977) recommended using the brewery-waste product as an essential source of vitamin B12 for Clarias gariepinus fingerlings. Tidwell et al. (1993) observed remarkable growth in pond-raised Macrobrachium rosenbergii at 40% distillers dried grains with solubles (DDGS) diets. Kaur (2004) observed that incorporation of brewery spent grain at 30% in supplementary fish feed, replacing rice bran, improved growth in Catla catla (Ham.), Labeo rohita (Ham.) and Cirrhinus mrigala (Ham.). The present study was there fore designed to investigate, the effect of the diets prepared with DDGW replacing GNOC on the growth performance and digestive enzyme activity without affecting feed utilization parameters of L. rohita.

MATERIALS AND METHODS

Diet and experimental design

Dried distiller’s grain waste (DDGW) was acquired from a local market and ground to fine powder in a blender, and used it in diet preparation (Table I, Supplementary Fig. 1A). The control diet (0% DDGW) was prepared using groundnut oil cake, wheat flour, rice bran, vitamins and minerals. Three iso-nitrogenous experimental diets [30% crude protein (CP)], were formulated with DDGW, by replacing groundnut oil cake (GNOC) at 30%, 40%, and 50% (T1, T2, and T3), respectively. The proximate analysis of basal and experimental diets was performed by the method outlined in Association of Official Analytical Chemists (Horwitz and Latimer, 2005) (Table I).

 

Table I. Composition of dried distillery grain waste of feed ingredients for the experimental diets (T1 – T3).

Ingredients	C	T1 (30%)	T2 (40%)	T3 (50%)
DDGW	-	22.5	30	37.5
GNOC	75	52.5	45	37.5
Wheat flour	4	4	4	4
DORB	20	20	20	20
Vitamin premix	0.5	0.5	0.5	0.5
Mineral premix	0.5	0.5	0.5	0.5
Crude protein (%)	29.57	31.48	30.57	32.10
Ether extract (%)	5.28	5.04	5.34	5.28
Total ash (%)	5.04	3.97	3.98	4.01
Moisture (%)	6.52	7.21	7.20	7.21
Energy (K cal g-1)	469.4	475.4	475.6	477.4

 

DDGW, Dried distiller’s grain waste; GNOC, Groundnut oil cake; DORB, De-oiled rice bran.

 

Feeding trials were conducted in hapas measuring 10m*3m*1m in size and installed in the earthen ponds covered with bird fencing nets (Supplementary Fig. 1B). All the treatment and control groups were conducted in duplicates. The experimental animals were stocked @ 100 per m3 each happa of size 0.04±0.03 g and were procured from the fish farm, Erode Bhavanisagar Centre for Sustainable Aquaculture, Bhavanisagar (Supplementary Fig. 1C). Until the beginning of the trials, rohu juvenile was fed with a basal diet. The feeding trial was conducted for 60 days. Fishes were fed @10% of body weight (BW) for the first 30 days, followed by 5% BW for another 30 days, twice a day at 10:00 h in the morning and at 16:00 h in the evening (Supplementary Fig. 1D) (Vinod and Basavaraja, 2010; Mmanda et al., 2020).

Water quality measurements

In all experimental groups, the nutrient quality of pond water was recorded every fortnight during the trial period. The water quality parameters i.e., water temperature, pH, dissolved oxygen (DO), ammonia (NH3), nitrite (NO2), nitrate (NO3), inorganic phosphate (PO4), free CO2, total hardness, total alkalinity, total suspended solids (TSS), total dissolved solids (TDS) and electrical conductivity (EC) were analyzed using standard procedures (APHA, 2005).

Bio-growth analysis

Growth parameters such as weight gain, weight gain percentage (WG%), specific growth rate (SGR), feed conversion ratio (FCR), feed efficient ratio (FER), protein efficient ratio (PER) and survival rate were determined using the following standard formula.

Proximate and amino acid analysis

At the end of the feeding trial, the fish from each replicate hapa were collected (n=8) to determine the whole-body composition and amino acid profile. The body composition was estimated following standard protocols (AOAC, 2010). Precisely 5g dry powder form of control and DDGW incorporated feed and fish samples were analysed for amino acid analysis at ATOZ Pharmaceuticals PVT.LTD., Chennai, Tamil Nadu. Using HPLC, LACHROM L-7000 and a ChromNAV software system from JASCO-HPLC analysis.

Determination of digestive enzyme activities

At the end of the experiment, ten fishes were collected from each replicate, and using a normal homogenizer, prepared a 5% tissue homogenate. The whole procedure was carried out in ice-cold condition. Homogenized samples were centrifuged at 5000 rpm for 10 min at 4°C. The supernatant was collected in a 5 ml tube and stored at -20°C for enzyme assay. Suitable dilution of the samples were done depending on the requirement. The Bradford method was used to estimate the total protein content of each tissue sample in enzyme assays (Bradford, 1976).

Amylase activity was estimated as the reducing sugars produced due to the action of gluco-amylase, and amylase on carbohydrates was determined using the di-nitro salicylic acid (DNS) method (Rick and Stegbauer, 1974). Protease activity was estimated by the casein digestion method (Drapeau, 1976). The lipase activity was determined by the titrimetric method according to the procedure described by Cherry and Crandell (1932). Aspartate amino transferase (AST) and alanine aminotransferase (ALT), lactate dehydrogenase (LDH), and malate dehydrogenase (MDH) activity were assayed with muscle tissue homogenates as described by Wooten (1964).

Statistical analysis

Statistical Package for Social Sciences (SPSS) version 25.0 (IBM Corp.) was used to test the differences between various treatments by one-way analysis of variance (ANOVA). Duncan’s multiple range test (p<0.05) was used to find the significant difference between treatments.

RESULTS AND DISCUSSION

Growth performances and nutrient utilization of rohu juvenile

Water quality parameters recorded during the experimental period were in the acceptable range for fish. Water quality parameters play an essential role in the biology and physiology of fish (Cho and Kaushik, 1990), as fishes are sensitive to change in water quality parameters. Optimum water quality assures maximum survival rate, nutrient utilization, and fish growth. In the present study, the selected ponds for the installation of hapa were free from sewage discharge or other anthropogenic activities. All the physicochemical parameters of water, such as water temperature (26.3-28.8oC), pH (7.2-7.8), dissolved oxygen (DO, 4.0-4.1 mgL-1), ammonia (NH3, 0.02-0.03 mg L-1), nitrite (NO2, 0.8-0.9 mg L-1), nitrate (NO3, 2.9-3.4 mg L-1), inorganic phosphate (PO4, 1.77-1.89 mg L-1), free (CO2, <0.01 mg L-1), total hardness (78-82 mg L-1), total alkalinity (77.2-81.0 mg L-1), total suspended solids (TSS, 312-322 mg L-1), total dissolved solids (TDS, 229-238 mg L-1) and electrical conductivity (EC, 0.28-0.33µ mhos cm-1) were within the optimum range of requirements for fish during the experimental period. Temperature is the main factor determining the growth and wellbeing of the fish. Das et al. (2005) found that the temperature ranges of 26–36 °C are not fatal to L. rohita juvenile. The optimum temperature range for growth was 25–28°C, temperature range during the feeding trial was within this recommended level. The water pH in the experimental pond ranged from 7.2-7.8, which is well within the acceptable range as suggested by Banerjea (1967). The ammonia values were in the range of 0.02-0.03 mg L-1. Jhingran (1991) suggested that the ammonia concentration of water must be in the range of 0.05-0.1 mg L-1. The present experimental study’s water quality parameters were in the optimal range (Harshavardhan et al., 2021).

Growth performance and nutrient utilization of rohu juvenile fed after 60 days was presented in Table II. An apparent increase in weight gain and improved feed utilization during the experimental period was observed. The nutritional quality of DDGW in rohu utilization from the feed was determined in terms of body weight gain (WG %), specific growth rate (SGR), feed conversion ratio (FCR), and protein efficiency ratio (PER). The growth and nutrient utilization parameters viz., WG (%), SGR, FCR, and PER are presented in Table II. The weight gain percentage, SGR, FCR, and PER vary significantly (p<0.05) among the various treatment groups. The results showed that the final weight and weight gain were significantly higher in fish fed at 50% DDGW diet group than in the other diet groups (p<0.05). Similarly, FCR was superior in the treatment group compared to the control group. The survival rates among the experimental groups did not vary significantly (p>0.05) (Table II). Maximum and minimum survival rates were found in T3 (54.02±0.21) and control (C) group (31.12±0.03), respectively among the different treatments.

 

Table II. Growth performance of rohu juvenile fed with DDGW during the experiment.

Treatments	0th day	15th day	30th day	45th day	60th day	Survival %
C	0.04 ± 0.02	0.29 ± 0.13	0.42 ± 0.02	0.84 ± 0.16c	1.53 ± 0.04c	31.12 ± 0.03
DDGW
T1 (30%)	0.04 ± 0.02	0.32 ± 0.19	0.46 ± 0.02	0.86 ± 0.32c	1.84 ± 0.07c	30.24 ± 0.02
T2 (40%)	0.04 ± 0.02	0.42 ± 0.10	0.73 ± 0.02	1.25 ± 0.06b	2.91 ± 0.02b	46.24 ± 0.15
T3 (50%)	0.04 ± 0.02	0.59 ± 0.13	0.87 ± 0.04	2.58 ± 0.16a	4.12 ± 0.02a	54.02 ± 0.21

 

Data expressed as mean ± standard error (M±SE); (n=15, r=2); Mean values in the same column with different superscript differ significantly (p<0.05). C, Control; T, Treatment; DDGW, Dried distiller’s grain waste.

 

Table III. Growth parameters of rohu juvenile fed with DDGW at different levels by replacement of GNOC.

Parameters	30th day				60th Day
	C	T1	T2	T3	C	T1	T2	T3
MIW	0.04± 0.02	0.04± 0.02	0.04± 0.02	0.04± 0.02	0.04± 0.02	0.04± 0.02	0.04 ± 0.02	0.04 ±0.02
MFW	0.42± 0.02	0.46± 0.02	0.73± 0.02	0.87± 0.04	1.53c ±0.04	1.84c ±0.07	2.91b ±0.02	4.12a ±0.02
MWG	0.38± 0.01	0.42± 0.02	0.69± 0.02	0.83± 0.04	1.49c ±0.02	1.80c ±0.03	2.87b ±0.02	4.08a ±0.03
WG%	950c ±138.05	1050c ±166.37	1725b ±166.37	2087a ±291.16	3725c ±291.16	4500c ±415.94	7175b ±166.37	10200a ±124.78
SGR	3.92b ±0.10	4.07b ±0.10	4.84ab ±0.06	5.14a ±0.09	6.07b ±0.05	6.38b ±0.06	7.15a ±0.06	7.72a ±0.08
FCR	1.33a ±0.01	1.26a ±0.06	1.13b ±0.01	1.08b ±0.09	0.97a ±0.01	0.87b ±0.05	0.85b ±0.01	0.79c ±0.01
FER	0.75b ±0.07	0.79b ±0.16	0.87a ±0.31	0.92a ±0.19	1.03c ±0.08	1.15b ±0.04	1.17b ±0.15	1.26a ±0.08
PER	1.27b ±0.65	1.29b ±0.29	1.37ab ±0.57	1.62a ±0.58	0.87b ±0.05	0.89b ±0.12	0.97a ±0.18	1.07a ±0.11
p value	>0.05	>0.05	>0.05	>0.05	>0.05	>0.05	>0.05	>0.05

 

Data expressed as mean ± standard error (M ± SE); n=2. Means in the same row having different superscripts are significantly different (p<0.05).

C, Control; T, Treatment; MIW, Mean Initial Weight; MFW, Mean Final Weight; MWG, Mean Weight Gain; SGR, Specific Growth Rate; FCR, Feed Conversion Ratio; FER, Feed Efficiency Ratio; PER, Protein Efficiency Ratio.

 

In the present study, there was a significant difference (p<0.05) in weight gain of L. rohita juvenile-fed diets that contained the graded levels of dried distiller grain waste (DDGW) with that of the control (C) diet and other experimental diets. Feeding dried brewery waste to livestock has been common practice since the initiation of beer production (Westendorf and Wohlt, 2002). The surplus availability of dried brewery waste makes it an excellent alternative to plant protein sources like soybean meal and GNOC in aqua-feeds. In the present investigation, inclusion at 50% DDGW diet in the T3 had the highest weight gain percentage (10200±124.78), SGR (7.72±0.08), FCR (0.79±0.01), and PER (1.07±0.11). Oliva-Teles and Goncalves (2001) reported that 50% of the brewery waste could replace fish meal with no adverse effect on the growth in sea bass. Similar results were reported in striped catfish fingerlings (Jayant et al., 2018). Zerai et al. (2008) reported that 50% brewery waste replacing fish meal promotes better growth and feed conversion in tilapia.

 

Table IV. Carcass composition of whole body of early rohu fingerlings of different experimental groups (% Wet weight basis).

Treatments	Moisture %	DM %	CP %	Lipid %	Ash %
C	76.22 ± 0.20	23.78 ± 0.20	14.12 ± 0.11	2.73 ± 0.13	3.51 ± 0.01
T1	75.78 ± 0.59	24.22 ± 0.59	15.88 ± 0.06	2.53 ± 0.05	3.43 ± 0.07
T2	75.00 ± 0.82	25.00 ± 0.82	16.56 ± 0.42	2.84 ± 0.21	3.56 ± 0.07
T3	73.49 ± 0.17	26.51 ± 0.17	17.05 ± 0.10	3.38 ± 0.12	3.61 ± 0.04

 

Data are expressed as mean ± standard error (M ± SE), n=2. C, Control; T, Treatment; DM, Dry matter; CP, Crude protein.

 

In channel catfish (Ictalurus punctatus), DDGW was used to replace fish and soybean meals, up to 40% of the diet, without lysine supplementation (Webster et al. 1991, 1992, 1993; Lim and Lee, 2009) and no negative repercussions on growth performance. Higher dietary replacement levels might be achieved with the adequate restoration of the dietary essential amino acid profile by using amino acid supplements or a combination among different protein sources (Webster et al., 1991; Cheng and Hardy, 2004). In channel catfish (Ictalurus punctatus), the dietary lysine supplementation allowed increased DDGW in the diet up to 70% (Webster et al., 1991; Robinson and Li, 2008). In blue catfish (Ictalurus furcatus), Webster et al. (1992a) stated that a combination of DDGW with soybean meal (35% DDGW and 49% soybean meal) could replace fish meal in the diet with or without lysine supplementation and methionine.

Carcass composition

The body composition of fish fed with different experimental diets is shown in Table IV. The highest body CP% was observed for fishes fed with the T3 diet, which was not significantly different than fishes fed with the other diets. The lowest body CP% was observed in fishes fed with the control diet. The highest tissue lipid accumulation was also recorded in the fish fed diet T3. The whole body moisture content (%) was lowest in the carcass of fish fed diet T3, and it was highest for the control diet and T1. The ash % did not vary significantly (p>0.05) among the groups fed different experimental diets.

Digestive and metabolic enzymes

Digestive and metabolic enzyme activities of L. rohita fed with different experimental diets were carried out after 60 days of the feeding trial. The activity of digestive enzymes like protease and amylase L. rohita juveniles’ intestine was found significantly (p<0.05) higher in the fish-fed diets containing among different experimental groups than the control and T1 fed fish (Table V). The maximum protease and amylase activities were noticed in the fish-fed diet T3, though it was not significantly (p<0.05) different from other diets of T1, T2 and control. However, the activities of the lipase enzyme were similar (p>0.05) among different experimental groups.

 

Table V. Specific enzyme activities in the digestive tract of early rohu fingerling fed with DDGW with different experimental group.

Treatments	Amylase	Protease	Lipase
C	3.65b ± 0.12	0.30b ± 0.57	0.68 ± 0.33
T1	3.96ab ± 0.09	0.34ab ± 0.37	0.67 ± 0.19
T2	4.13a ± 0.10	0.34ab ± 0.25	0.70 ± 0.21
T3	4.69a ± 1.17	0.39a ± 0.21	0.69 ± 0.20
p value	<0.05	<0.05	>0.05

 

Data are expressed as mean ± standard error (M ± SE). (n=15, r=3); Mean values in the same column having different superscripts differ significantly (p<0.05). C, Control; T, Treatment. Protease as micromol of tyrosine released/ min/mg protein. Amylase as micromol of maltose released/min/mg protein. Lipase as units/mg protein.

 

The metabolic enzyme activities of L. rohita fed with different experimental diets were shown in Table V. The Aspartate amino transaminase (AST) and alanine amino transaminase (ALT) activity in the muscle T3 fed groups was statistically higher (p<0.05) than that of the control and other groups. The lactate dehydrogenase (LDH) and malate dehydrogenase (MDH) activity in the muscle of the fish fed different experimental diets did not show any significant variation (p>0.05). The activities of digestive enzymes (protease, lipase, and amylase) were higher in T3 (50% DDGW), followed by T2 and T1 experimental groups. In contrast, the lowest activity of these enzymes was observed in the control (C) group. There was no significant difference between the experimental groups included with DDGW. Similar findings by Lim and Yildirim (2008) was suggested that lysine supplementation at 40% of DDGW in the diet has shown better growth performance, feed utilization, and survival of juvenile channel catfish. In the present study, metabolic enzyme activities like AST, ALT, LDH, and MDH have shown no significant difference between the treatments indicating low stress due to the experimental feed (Chatterjee et al., 2003).

 

Table VI. Metabolic enzyme activities of muscle of L. rohita fed with different replacement level of DDGW with the GNOC.

Treatments	AST Muscle	ALT Muscle	LDH Muscle	MDH Muscle
C	26.62c±0.13	14.84c±0.70	4.09±0.22	3.01±0.01
T1	27.31bc±0.46	15.14c±0.41	3.97±0.04	2.71±0.02
T2	28.09b±0.13	16.86b±0.26	3.98±0.03	2.48±0.03
T3	29.99a±0.96	17.43a±0.41	3.62±0.02	2.34±0.02

 

Data expressed as mean ± Standard Error (M±SE) (n=15, r=2); Mean values in the same column with different superscript differ significantly (p<0.05). C, Control; T, Treatment. ALT, specific activities expressed as Nano moles of sodium pyruvate formed/mg protein/minute at 370C. AST, specific activities expressed as Nano moles of oxaloacetate released/min/mg protein at 370C. LDH, specific activities expressed as units/mg protein/ min at 37°C; MDH, specific activities expressed as units/mg protein/ min at 37°C.

 

Amino acid analysis

The amino acids profile of DDGW of feed and fish is presented in Tables VII and VIII. Essential amino acids like histidine, threonine and methionine were higher in the T3 fish group supplemented with the 50% DDGW than control feed. The essential aminoacids were found higher in DDGW incorporated diets than that of control diet. Webster et al. (1992) revealed that growth rate and feed conversion indices were not affected by the complete substitution of soybean meal with distiller’s grain soluble (DGS) in the diet of channel catfish. Similarly, Jayant et al. (2018) also reported that protein intake values were significantly (p<0.05) higher in fish fed at 25% and 50% brewery spent grain (BSG) levels in striped catfish (Pangasianodon hypophthalmus) fingerlings. No adversegrowth effect reportd in yellow perch (Perca flavescens), fed with

 

Table VII. Amino acid profile of DDGW of feed.

Amino acid (mg/5g DM)	C Feed	T1 Feed	T2 Feed	T3 Feed
Essential amino acids
Arginine	18.08	19.01	19.6	20.0
Histidine	8.9	9.4	9.6	10.2
Isoleucine	13.8	13.5	13.7	13.9
Leucine	26.8	25.2	25.6	26.0
Lysine	17.4	16.8	16.4	15.8
Phenylalanine	14.6	14.4	14.3	14.4
Methionine	9.6	9.3	9.1	8.9
Threonine	11.1	11.3	11.6	11.7
Tryptophan	2.9	2.9	2.8	2.6
Valine	14.3	14.5	14.6	14.7
Non-essential amino acids
Alanine	18.3	16.1	15.8	15.2
Glycine	22.1	21.6	21.0	20.8
Aspartic acid	33.4	32.8	32.1	31.3
Glutamic acid	62.4	64.3	64.9	65.4
Serine	13.7	14.6	15.4	15.9
Tyrosine	10.4	11.5	11.9	13.01

 

DM, Dry Matter; C, Control; T, Treatment.

 

Table VIII. Amino acid Profile of DDGW of fish.

Amino Acid (mg/5g DM)	C Fish	T1 Fish	T2 Fish	T3 Fish
Essential amino acids
Arginine	11.1	12.5	12.4	12.7
Histidine	5.9	6.7	6.6	6.4
Isoleucine	8.9	9.3	9.4	9.4
Leucine	10.9	13.1	13.4	13.7
Lysine	11.8	12.2	12.6	12.9
Phenylalanine	6.8	7.8	7.6	7.1
Methionine	4.9	5.8	5.9	6.1
Threonine	8.3	9.4	9.6	9.6
Tryptophan	2.5	2.6	2.7	2.7
Valine	8.3	9.2	9.3	9.3
Non-essential amino acids
Alanine	15.1	15.5	15.3	15.2
Glycine	13.4	13.5	13.6	13.7
Aspartic acid	25.9	26.1	26.0	26.2
Glutamic acid	27.9	27.8	28.3	28.4
Serine	4.2	6.2	6.4	6.7
Tyrosine	3.1	3.7	3.7	3.4

 

DM, Dry Mass; C, Control; T, Treatment.

 

DDGW and soybean meal mixture up to 49.5% (Schaeffer et al., 2011). The improved growth performance of rohu juvenile in the present study suggests that protein and nucleic acid content in DDGW improved the nutrient utilization and growth performance of L. rohita juvenile. In the present study, the amino acid concentrations like lysine, leucine, threonine, and methionine contents in DDGW fishes showed higher, in fish fed with 50% DDGW than the T2, T3, and control group. Zerai et al. (2008) had seen similar increases of essential amino acids in the 25% to 50% inclusion levels of brewery waste with partial replacement of fish meal in Nile tilapia diet. There are some hindrances in using GNOC as fish feed ingredient. Although it is an excellent source of the amino acid arginine, it is deficient in sulphur containing amino acids like lysine, cysteine and methionine (Green et al., 1988).

CONCLUSION

The rising cost and the limited supply of conventional fish feed ingredients have necessitated focusing research efforts towards the potential utilisation of energy and proteins from several agribyproducts that are cheaper with high nutritive value. During the study period, traditionally local farmers generally use a 50:50 mixture of rice bran (@ 20/kg) and GNOC (@ 45/kg) which would cost Rs. 32 per kg, while when DDGW (@ 27/kg) when used the cost of the feed would be around Rs. 28/- per kg. The present study showed improved growth performance, nutrient utilization, and survival rate of juvenile L. rohita fed with a diet of 50% DDGW. Hence, it can be conluded that utilization of DDGW upto 50% replacement of GNOC can be scientifically and economically better than the traditional practice of utilizing GNOC and rice bran alone in the rearing of L. rohita in the nursery phase.

Acknowledgement

The present study was an outcome of the student research carried out for the award of Ph.D. The study was supported by Tamil Nadu Dr. J. Jayalalithaa Fisheries University, Nagapattinam, Tamil Nadu, India, in the form of a fellowship. We also thank the Director, Directorate of Sustainable Aquaculture, TNJFU, for necessary infrastructure facilities as well as scientific support for carrying out the research work.

Funding

The study received no external funds.

IRB approval

The research committee of the University has approved the work.

Ethical statement

The experiment was conducted following the procedures 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 care and use of animals in scientific research. The procedural handling and rearing of the fish were performed according to the animal ethics committee, Tamil Nadu Dr. J. Jayalalithaa Fisheries University, Nagapattinam.

Data availability statement

The data that support the findings of this study are available within the article.

Supplementary material

There is supplementary material associated with this article. Access the material online at: https://dx.doi.org/10.17582/journal.pjz/20230514060530

Statement of conflict of interest

The authors have declared no conflict of interest.

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