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Evaluation of Locally Available Plant Meals as a Replacement of Soybean Meal as Dietary Protein Source for Catla catla Fingerlings.

Byline: Rafia Tayyab, Noor Khan, Muhammad Sharif Mughal, Naureen Aziz Qureshi, Muhammad Naeem Khan and Fayyaz Rasool

Abstract The possibility of locally available plant meals as a replacement of soybean meal as dietary protein source for Catla catla fingerlings was evaluated under intensive rearing condition in glass aquarium for 60 days.

Three iso-nitrogenous (24% crude protein) diets (D1, D2 and D3) were prepared and fed to fish at 3% wet body weight per day. Each treatment group had two replicates. The data was analyzed by employing one way ANOVA. The survival rate of fish as observed in D1 and D2 was 75%, while it was 100% in D3.

The study showed across the board poor fish growth in all the treatment groups with non-significant difference (P [greater than or equal to] 0.05) in final weight, gain in weight, SGR and FCR. However, a significant difference (P Less than 0.05) was observed for fish whole body crude protein, crude lipid, ash and crude fiber contents. Crude protein and ash contents significantly increased with the increased level of plant protein meals while crude lipid percentage decreased with increasing the plant meals.

It can be concluded that this replacement of soybean meal with other local plant protein meals did not have significant effect on fish growth. However, inclusion of higher level of plant meals (Ipil ipil leaf meals, sunflower meals and rice polish) resulted in increased crude protein, fiber and ash contents of the fish.

Key Words: Plant source meals, Catla catla, growth, proximate composition.


Feed and nutrition is critical in fish farming because feed represent over 60% of the production costs (Li and Wang, 2004). Presently, aquaculture production is greatly constrained by undersupply, scarcity and high cost of conventional fish feed.

The ever growing cost and uncertainties

about the quality and availability of fish and soybean meal have compelled the aquaculture nutritionists and feed manufactures to search and replace this with cheap and readily available plant protein sources.

Besides cost, improvement in feed and nutrition of aquatic species is also necessary by enhancing the quantity and nutritional quality of fish flesh (Khan et al., 2011). Identification of suitable alternate protein sources for inclusion in fish feeds becomes imperative to counter the scarcity of fish meal and soybean meal.

A variety of plant protein sources including sunflower meal, ipil ipil leaf meal

(Amisah et al., 2009; Bairagi et al., 2004), cotton seed meal, raw soybean meal (Patnaik et al., 2005) and extract of Moringa leaves (Afuang et al., 2003; Ritcher et al., 2003) have been tested and included as alternatives to fish meal.

Since sunflower meal contains about 38% crude protein (Shazadi et al.,

2006), rice polish 12-16% (Abid and Ahmed, 2009)

and Leucaena leaf about 22.7%, these can be good candidate to replace as cheaper and quality ingredient in aqua feed (Atawodi et al., 2008). The nutritive value and digestibility of Ipil ipil is higher than that of tropical grasses (Zamal et al., 2009).

Pakistan, an agro-based country has the potential to sustain the production of various plants based feed ingredients (rice polish, sunflower meal and Ipil ipil, etc) that can be utilized to replace costly protein ingredients such as fish and soybean meals to produce cost effective aqua feeds.

Recently few commercial aqua feed industries are utilizing costly imported soybean meal and incorporating them in the commercial fish feed.

As already mentioned Pakistan has plenty of plant based feed ingredients that are sustainable and cost effective and can be utilized for our commercially important fish species. Catla catla is one of the three most popular commercially important culturable herbivorous fish in South Asia including Pakistan. It is a fast growing and surface feeding fish and can utilize efficiently mash feed from the water surface as well.

The present study was therefore planned to study the effect of locally available plant based feed ingredients diet with the replacement of soybean meal diet for the survival, growth and body composition of Catla catla fingerlings.


The study was carried out at Research and Training Facilities, Department of Fisheries and Aquaculture, University of Veterinary and Animal Sciences, Ravi Campus, Pattoki, Pakistan under intensive rearing conditions. Fingerlings of Catla catla (average weight 8.0 g) were procured from Fisheries Research and Training Institute, Manawan, Lahore and kept in glass aquaria (60.96 x

55.88 x 40.64 cm) by using two replicates of each

diet for 60 days.

Fortnightly morphometric measurements (Total Body Weight and Total Body Length) of fish were conducted to monitor the growth and adjusting the daily ration requirement. Growth indices such as net weight gain (NWG), feed conversion ratio (FCR), and specific growth rate (SGR) were calculated by using following formulae:

NWG = Final body weight (g) - Initial body weight (g) FCR =Feed intake (g) / Wet weight gain

SGR %/day = ln(W1 )- ln(W2 ) x100 /No. of days

Artificial feed formulation

Three types of experimental mash feeds (D1,

D2 and D3) were prepared by using fish meal, soybean meal, ipil ipil leaf meal, rice polish, wheat bran, sunflower meal and Nutrimix as feed ingredients (Table I).

Experimental feeds such as

0% (D1), 20% (D2) and 40% (D3) soybean meal was replaced with plant based ingredients meals like Ipil ipil leaf meal, sunflower meal and rice polish. The fish was fed twice a day at 3% fish wet body weight up for 60 days.

Chemical analysis

The formulated feeds, feed ingredients and fish samples were analyzed for dry matter, ash, crude protein, crude lipid and gross energy as per AOAC (2006).

Water quality parameters

Water quality parameters viz., temperature, dissolved oxygen (DO), total dissolved solids (TDS) and pH were monitored on daily basis, while nitrates were determined on fortnightly basis by using spectrophotometer (APHA, 1998).

Statistical analysis

The data collected after completion of trials were statistically analyzed using Analysis of Variance (ANOVA) (Steel et al., 1997) using Minitab Statistical package 1.5. Comparison of means was done by using Tuky's t-test. The significance of probability was tested at PLess than0.05 level.


Growth parameters of the current study showed non-significant differences (p [greater than or equal to] 0.05) in final weight gain and net weight gain in all the treatment groups (Table II). The SGR was 0.62%,

0.36% to 0.66%/day in D1, D2 and D3, respectively also revealed non-significantly poor growth performance. There were non-significantly (p [greater than or equal to]

0.05) higher values of feed conversion ratio (FCR) D1 4.71, D2 7.86 and D3 4.79 (Table II).

Proximate composition of whole body

revealed significant differences (p Less than 0.05) among different treatment groups like; ash contents were significantly higher in D1 and D2 compared to D3 (Table III). Crude protein percentage was found significantly higher in D3 than D1 and D2. Crude lipid contents were found significantly higher in D1 followed by D2 and D3, respectively. Crude fiber was significantly higher in D2 followed by D1 and D3 (Table III).

The results of physico-chemical parameters

such as temperature ranged from (28-30degC), dissolved oxygen (2.4-3.58 mg/l), pH (7.98-8.16), salinity (0.85-0.91ppt), total dissolved solids (1531.83-1646.99us/cm) and nitrates (0.00017mg/l).

Table I.- Ingredients composition and crude protein (CP) concentration of the experimental diets

Feed ingredients D1 (0% CP% D2 (20% CP% D3 (40% CP%

soybean meal) soybean meal) soybean meal)

Fish meal 10 5.54 10 5.54 10 5.54

Soybean meal 20 8.05 10 4.03 0 0.0

Ipil ipil leaf meal 0 0 20 4.6 40 9.2

Rice polish 45 6.4 30 4.27 15 2.13

Wheat bran 25 4.15 23 3.81 21 3.48

Sunflower meal 0 0 7 1.86 14 3.71

Total 100 24.14 100 24.11 100 24.06

CP, crude protein % in diet.

Table II.- Comparison of growth parameters of Catla catla fingerlings fed 24% crude protein diets (means are of two replicates n = 2).

Parameters D1 (0% soybean D2 (20% soybean D3 (40% soybean PSEM ANOVA (P value)

meal) meal) meal)

No. of fish stocked 8 8 8 - -

Survival % 75 75 100 - -

Initial wt (g) 8.03a 8.22a 9.46a 3.52 0.910

Final wt (g) 11.48a 11.24a 13.58a 4.66 0.865

Net wt. gain (g) 3.46a 3.02a 4.13a 1.18 0.674

SGR% 0.62a 0.36a 0.66a 0.12 0.476

FCR 4.71a 7.86a 4.79a 1.8 0.283

Figures with same letters are not significantly different (P Greater than 0.05); PSEM, pooled standard error of mean.

Table III.- Proximate composition of Catla catla fingerlings fed 24% crude protein diets

Days D1 (0% soybean D2 (20% soybean D3 (40% soybean PSEM ANOVA (P value)

meal) meal) meal)

DM (%) 99.0a 98.75a 99.39a 0.495 0.510

Ash (%) 12.07a 10.61a 9.46b 0.522 0.035

CP (%) 64.04a 66.18a 70.0b 0.77 0.010

CL (%) 15.03a 12.39b 8.03c 0.370 0.001

NFE (%) 1.86a 2.58b 1.25c 0.12 0.003

Figures with different superscripts are significantly different (P Less than 0.05); DM, dry matter; CP, crude protein; CL, crude lipid; NFE, nitrogen free extract; PSEM, Pooled standard error of mean.

Non-significant difference (p [greater than or equal to] 0.05) was observed for all the parameters among treatments except dissolved oxygen which revealed significant differences between D1 and D3 (Table IV).


In present study growth parameters showed non-significant differences in final weight gain and net weight gain in all the treatment groups. Some

mortality (25%) was observed in D1 and D2 while no mortality (0.0 %) was observed in D3 where soybean meal was replaced with 40% ipil-ipil leaf meal, 15% rice polish and 14% sunflower meal.

The possible reason behind mortality and poor fish growth are not clear but can be related to low dissolved oxygen concentration in the aquarium which ranged from 2.4 to 3.5 mg/L.

The SGR values during current study were 0.62%, 0.36% to

0.66%/day in D1, D2 and D3 revealed poor growth

Table IV.- Physicochemical parameters of the experimental trial of Catla catla fingerlings fed 24% crude protein diets

(means are of two replicates n = 2).

Parameters D1 (0% soybean D2 (20% soybean D3 (40% soybean PSEM ANOVA (P value)

meal) meal) meal)

Temp. (oC) 28.83a 30.01a 30.1a 1.41 0.388

DO (mg/l) 3.58a 2.72ab 2.44b 0.48 0.021

pH 8.16a 7.98a 8.01a 0.10 0.076

Salinity (ppt) 0.85a 0.91a 0.88a 0.12 0.800

TDS (us/cm) 1646.997a 1531.83a 1563.224a 160.1 0.594

Nitrates (mg/l) 0.00017a 0.00017a 0.00017a 0.00 0.866

Figures with different superscripts are significantly different (P Less than 0.05); DO, dissolved oxygen; TDS, total dissolved solids; PSEM, pooled standard error of mean.

performance. Similarly, the FCR values were also found poor and non-significantly different in D1 at

4.71, D2 at 7.86 and D3 at 4.79. Contrary to our results, Zamal et al. (2009) reported that 15% level of ipil-ipil in the diets have good nutritive values and have a significant effect on the growth, FCR and FCE of Oreochromis niloticus.

Their findings revealed that ipil-ipil leaf meal could be used as protein substitute up to 25% and optimum level 15% in diet of growing tilapia. Um-e-Kalsoom et al. (2009) reported feed conversion ratio (FCR) of blood meal (1.68 +- 0.96), followed by rice broken (1.64 +- 0.64) and wheat bran (1.59 +- 0.71) in hybrid fish (Catla catla x Labeo rohita) which are much lower and better than present study.

Abid and Ahmed (2009) reported SGR 3.7% and FCR 1.5 for rohu fingerlings while feeding 100% rice polish. The possible reason of this contradiction in results of higher FCR in current study might be due to the differences in rearing conditions such as glass aquaria, low oxygen (stress), feed preparation, feed intake and inclusion of other plant based meals such as ipil ipil, sunflower meal, wheat bran etc.

De Silva and Davy (1992) reported that feed digestibility plays an important role in lowering the FCR by efficient feed utilization.

In the present study, diet (D3) containing

40% Ipil ipil leaf meal, 15% rice polish and 14%

sunflower meal comparatively showed slightly higher growth when compared with D2 and D1.

100% survival in D3 might be due to some growth

promoting agents in plant source meals that may have enhanced the survival and growth of fingerlings in hypoxic condition. Hassan et al.

(1994) has reported a trend of reduced growth performance of Indian major carps fed with diets containing 25% soaked Leucaena diets, while Ghatnekar et al. (1983), by using 30-65% Leucaena leucocephala recorded no adverse effects on growth and even 15% level used in the diets have good nutritive values and played a significant role on the growth, FCR and FCE in Oreochromis niloticus.

The results of proximate composition of fish whole body revealed that increase in the level of both ipil-ipil leaf meal and sunflower meal increased the protein contents significantly. However, Crude lipid and ash contents during this study showed opposite trend.

According to Zamal et al. (2009) Ipil ipil and leaf material of Leucaena is a good source of crude protein (CP), calcium (Ca) and phosphorus (P), and carotene that might enhanced the protein contents in the experimental fish.

The range of physico-chemical parameters

were found within acceptable limit (Rahman et al.,

2009) except dissolved oxygen for Catla catla fingerlings. The reason of such differences is not clear but that might be associated with higher fish growth and metabolic rate in treatment group D3 where fish showed numerically higher growth compared to D1.


It can be concluded from the present study that replacement of soybean meal with locally available plant meals (Ipil ipil leaf meal, sunflower meal and rice polish) did not significantly increase the growth of fish.

However, higher level of inclusion of Ipil ipil leaf meal and sunflower meal resulted in higher survival rate, increased crude protein and crude fiber contents.

It can be suggested that ipil ipil whole plant meal may be incorporated instead of leaf meal with other local plant origin protein sources to prepare pelleted feed that binds the ingredients firmly and may be applied in field condition for long term basis (6-9 months). Additionally detailed chemical analysis of ipil ipil may also be done to explore the growth inhibiting factors for future research.


The authors are extremely thankful to the Chairman, Department of Fisheries and Aquaculture, University of Veterinary and Animal Sciences, Lahore, Pakistan for providing facilities for conducting this study in Fish Hatchery aquaria.


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