Livestock Research for Rural Development 21 (3) 2009 | Guide for preparation of papers | LRRD News | Citation of this paper |
To study the daily excretion patterns of wastes like ammonical nitrogen (NH4-N) and ortho-phosphate (o-PO4) fish were fed on 10 iso nitrogenous diets (D1 to D10) formulated by replacing fishmeal (FM) with defatted canola at 4 inclusion levels (25, 50, 75 and 100g/kg) with and without supplementing the diets with a mineral premix and amino acids (MPA).
Studies have reveled that oxygen levels (DO) fluctuated between 4 to 5 mg/l and pH remained alkaline (7.5 to 7.9). Significantly (p<0.05) highest conc. of NH4-N and o-PO4 were observed in the water medium in which fish were fed on reference diets D1 and D6 containing FM as the main protein source. The excretion decreased on increasing the inclusion levels of defatted canola. Further, in the groups of fish fed on diets D6 to D10 the excretion of NH4-N and o-PO4 was lower than those observed in groups of fish where diets D1 to D5 were used indicating that incorporation of MPA reduces the excretion of NH4-N and o-PO4 in water medium. The peak time of excretion of NH4-N was observed at the end of 6 hrs. of post-feeding and of o-PO4 was observed at the end of 8 hrs. of post-feeding.
Key words: Ammonical nitrogen, Channa punctatus, defatted canola, excretion, fishmeal, ortho-phosphate
Proteins are the main source of nitrogen and essential amino acids and also the most expensive energy source (Pillay 1992).To maximize the nutrient utilization and minimize the solid and soluble waste load, it is essential to provide cultured fish with optimum level of protein (Cho 1993). Generally, nutrients absorbed in excess of requirement may be excreted as ammonia and urea (Beveridge and Phillips 1993). When food wastage is high and nitrogen retention and assimilation are poor, a major portion of nitrogen is added to the culture system, which may ultimately pollute the environment (Handy and Paxton 1993).
Fishmeal, which is considered as the best source of protein for fish is difficult to get in interior parts and even if it is available, it will be very costly To reduce feed cost and substitute of minerals and protein components of diets, materials of plant origin such as soybean, canola (rapeseed), cottonseed meal etc. to varying degrees were suggested by Hastings (1976), Jackson et al (1982), Viola et al (1982, 1983); Lall (1991 ), Kaushik (1992); Vielman et al (2000), Jindal (2001), Priyanka and Garg (2002), Jindal and Garg (2005), Robinson and Menghe (2007).
Rapeseed and mustard oil is primarily used for edible purposes, while the defatted meal is utilized as animal feed. Black and dark brown seed coat colour is of normal occurrence in rapeseed mustard. Further, canola (Brassica napus) is currently defined as having less than 2% erusic acid in the oil and less than 30 micromoles of the aliphatic glucosinolates per gram of oil free meal. Canola meal, has about 40% protein (on a dry matter, oil free basis) and a relatively well balanced amino-acid composition. Canola meal is a high protein feed ingredient of plant origin, however, it possesses some anti-nutritional compounds such as glycosides and tannins. Furthermore, protein in canola is surrounded by relatively indigestible carbohydrate, which cannot be broken down without the use of added enzymes (Buchman et al 1997).
Fish excrete phosphorus in soluble and particulate forms (Lall 1979, 1991 and Pillay 1992; Vielman et al 2000) The soluble fraction is called ortho phosphate (o-PO4), is most available for plant growth (Bostrom et al 1988). However the main loading of phosphorus to the environment was reported to be via faecal pellets (Pillay 1992, Kibria et al 1996, 1998).
The main product excreted by teleosts fish is total ammonia nitrogen (TAN), which is formed in the lever and excreted across the gills .About 80-90% of nitrogen loss from fish is through gill excretion and the faecal nitrogen loss accounts for 10-20% .Nitrogen is also lost through uneaten feed or dust (Kibria et al 1996, 1998).(Figure 1).
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Therefore, there is a need to search alternate protein sources, that may also reduces the input of nitrogen and phosphorus into the environment.
Specimens of Channa punctatus were obtained from fish dealers of Hisar. Specimens with mean body weight (8.0 to 15.0 g) were used in the studies. Fish were placed in the transparent glass aquaria (60X30X30 cm) kept in the laboratory where the temperature was maintained at 25±10C and the lighting scheduled at 12h of light alternating with 12h of darkness. The fish were acclimatized for a minimum of 7 days prior to the initiation of experimental treatments. The water was renewed daily with chlorine free water.
Canola was used as the main protein source. It was defattened to remove antinutritional compounds prior to the preparation of experimental diets (Jindal 2001, Garg et al 2002).
Groundnut oil cake, Rice bran, fishmeal and defatted canola were finely ground to pass through 0.5 mm sieve. All the ingredients were mixed according to Table 1 and dough was made using distilled water.
Table 1. Ingredient Composition (%) of compounded diets from D1 to D10 |
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Ingredients |
Diet number |
||||||||||||||||
D1 |
D2 |
D3 |
D4 |
D5 |
D6 |
D7 |
D8 |
D9 |
D10 |
||||||||
|
60.0 |
60.0 |
60.0 |
60.0 |
60.0 |
60.0 |
60.0 |
60.0 |
60.0 |
60.0 |
|||||||
Rice brana2 |
24.0 |
24.0 |
24.0 |
24.0 |
24.0 |
23.0 |
23.0 |
23.0 |
23.0 |
23.0 |
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Fish meal (FM) |
10.0 |
7.5 |
5.0 |
2.5 |
- |
10.0 |
7.5 |
5.0 |
2.5 |
- |
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Defatted canola |
- |
2.5 |
5.0 |
7.5 |
10.0 |
- |
2.5 |
5.0 |
7.5 |
10.0 |
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Chromic oxide (Cr2O3) |
1.0 |
1.0 |
1.0 |
1.0 |
1.0 |
1.0 |
1.0 |
1.0 |
1.0 |
1.0 |
|||||||
Binder (Carboxyl methyl cellulose) |
5.0 |
5.0 |
5.0 |
5.0 |
5.0 |
5.0 |
5.0 |
5.0 |
5.0 |
5.0 |
|||||||
Mineral mix |
- |
- |
- |
- |
- |
1.0 |
1.0 |
1.0 |
1.0 |
1.0 |
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D1 and D6 : reference diets containing 100% fishmeal (without and with mineral mix) FM100, FM-M. D2 to D5 : containing fishmeal and defatted canola at various inclusion levels (25-100%)DC25, DC50, DC75, DC100.
D7 to D10
: containing fishmeal and defatted canola at various
inclusion levels (25-100%) supplemented with mineral mix a1 and a2 à used as basal feed ingredients; b and c used as main protein source. FM was replaced by defatted canola at each inclusion level. d used as an external indigestible marker for estimating apparent digestibility e used as binder to make the diets water stable f added to supplement the diets with minerals and amino acids.
Each Kg contains
Copper - 312mg; Cobalt - 45mg; Magnesium - 2.114g; Iron
-979mg; Zinc - 2.13g; Iodine 156mg; |
Thereafter, the dough was passed to a mechanical palletizer to obtain pellets (0.5 mm thick) which were dried in an oven and used in the studies for 45 days. The proximate composition of all the 10 diets is presented in Table 2.
Table 2. Proximate composition ( % dry weight ) of various experimental diets from D1 to D10 (FM 100 to DC100-AAM) |
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Diet No. |
Crude Protein |
Crude Fat |
Crude Fiber |
Total Ash |
Nitrogen Free Extract |
Gross Energy, KJg-1 |
FM100 |
33.83 |
6.66 |
9.30 |
12.66 |
37.53 |
17.07 |
DC25 |
31.79 |
6.63 |
9.43 |
12.23 |
39.77 |
16.95 |
DC50 |
31.50 |
6.53 |
10.36 |
10.50 |
41.10 |
17.07 |
DC75 |
30.92 |
6.50 |
10.30 |
10.40 |
41.88 |
17.06 |
DC100 |
30.63 |
6.30 |
10.60 |
9.76 |
42.71 |
17.05 |
FM-M |
33.83 |
6.70 |
9.36 |
12.70 |
37.40 |
17.06 |
DC25-AAM |
33.54 |
6.60 |
9.60 |
12.16 |
38.09 |
17.07 |
DC50-AAM |
32.38 |
6.56 |
10.36 |
11.26 |
39.43 |
17.01 |
DC75-AAM |
31.79 |
6.53 |
10.33 |
10.20 |
41.14 |
17.15 |
DC100-AAM |
31.50 |
6.36 |
10.56 |
9.16 |
42.40 |
17.23 |
Mean with same letter in the same column are not significantly (p>0.05) different |
On the last day of experiment offer the same feed to the fish in sufficient quantity so that the same is consumed, wait for 2 hours. Maintain a fixed level of water in each aquarium (say 30-40 L). Remove the excess of feed. Start collecting water samples from each aquarium in replicate of 2 for the determination of ammonical nitrogen (NH4-N) and ortho-phosphate (o-PO4) following APHA (1998) to see the influence of compounded feeds on pollution status of receiving water in the aquaria.
Calculate the excretory levels of NH4-N and o-PO4 in treated water as follows:
NH4-N excretion = NH4-N (mg l-1) in aquarium water
(mg/100g BW of fish) Fish weight (mg) per L of water
o-PO4 excretion = o-PO4 (mg l-1) in aquarium water
(mg/100g BW of fish) Fish weight (mg) per L of water
The oxygen levels (DO) fluctuated between 4-5 mg/l and pH remained alkaline (7.5 to 7.9). Significantly (p< 0.05) highest conc. of NH4-N and o-PO4 were observed in the water medium in which fish were fed on reference diets FM100 and FM-M containing FM as the main protein sources (Table 3).
The excretion decreased on increasing the inclusion levels of defatted canola.
Table 3. Water quality parameters of different aquariums stocked with Channa punctatus fingerlings fed on diets D1 (FM100) to D10 DC100-AAM containing defatted canola as the main protein source |
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Parameters |
Diet No. |
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FM100 |
DC25 |
DC50 |
DC75 |
DC100 |
FM-M |
DC25-AAM |
DC50-AAM |
DC75-AAM |
DC100-AAM |
|
Dissolved oxygen (DO), mg/l |
4.7 |
4.9 |
5.0 |
4.6 |
4.3 |
4.4 |
4.7 |
4.5 |
4.8 |
5.0 |
pH |
7.5 |
7.8 |
7.4 |
7.9 |
7.6 |
7.5 |
7.6 |
7.9 |
7.7 |
7.6 |
Water temperature, °C |
25.6 |
26.7 |
27.0 |
26.6 |
27.1 |
25.2 |
26.3 |
27.0 |
25.8 |
24.9 |
Conductivity micro (µ) mhos cm-1 |
0.47 |
0.50 |
0.52 |
0.51 |
0.48 |
0.47 |
0.53 |
0.55 |
0.58 |
0.57 |
Free carbon dioxide (Free CO2), mg/l |
17.3 |
17.8 |
16.5 |
16.3 |
17.2 |
17.3 |
17.4 |
17.9 |
17.0 |
17.3 |
Total alkalinity, mg/l |
254.0 |
250.0 |
248.0 |
242.0 |
253.0 |
247.0 |
241.0 |
251.0 |
245.0 |
252.0 |
Total hardness, mg/l |
227.0 |
225.0 |
229.0 |
241.0 |
235.0 |
231.0 |
235.0 |
240.0 |
228.0 |
225.0 |
Ammonical nitrogen (NH4-N) excretion, mg/100BW of fish |
0.703 |
0.620 |
0.570 |
0.516 |
0.536 |
0.650 |
0.613 |
0.540 |
0.523 |
0.480 |
Ortho-phosphate (o-PO4) excretion, mg/100BW of fish |
0.266 |
0.246 |
0.240 |
0.186 |
0.160 |
0.260 |
0.226 |
0.173 |
0.163 |
0.110 |
This is because the fish can digest plant proteins much more easier than animal proteins (Deepak and Garg 2003, Priyanka and Garg 2002, Jindal 2001, Kalla and Garg 2004)
Further in the groups of fish fed on diets FM-M to DC100-AAM, the excretion of NH4-N and o-PO4 was lower than those observed in groups of fish where diets FM100 to DC100 were used indicating that incorporation of MPA reduces the excretion of NH4-N and o-PO4 in water medium (Table -3). These results are in agreement with those of Viola and Lahav (1993). According to them the calculated amounts of excreted (not retained) nitrogen per kg gain was reduced by 20% in the lysine supplemented feeds, as compared to the 30% protein feed. Concomitantly, calculated phosphorus excretion per kg gain was also decreased approximately by 100%.
Water samples were analysed for 16 hrs at 2 hr interval of post-feeding revealed peaks in NH4-N and o-PO4 excretion (Figure 2 and Figure 3).
The peak time of excretion of NH4-N in groups of fish fed on diets FM100, DC25 and DC50 were observed at the end of 8 hrs. of post feeding but in groups of fish fed on diet DC75 and DC100 the peak time of excretion of NH4-N was slightly earlier i.e. at the end of 6 hrs. of post-feeding (Figure 2 A).
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Figure 2.
Excretion patterns of ammoniacal nitrogen (mg/100g body weight)
in treated waters in fish Channa punctatus fed on diets
D1 to D10 “A” without mineral mix and amino-acids (D1 to D5) “B” with mineral mix and amino-acids (D6 to D10) |
In the groups of fish fed on diets FM-M to DC100-AAM supplemented with MPA, the excretion of NH4-N was lower than those observed in groups of fish fed on diets FM100 to DC100 But the pattern of excretion of NH4-N was same as for groups of fish fed on diets FM100 to DC100 (Figure 2 B).
The peak time of excretion of o-PO4 in the groups of fish fed on the diets FM100 to DC100 were observed at the end of 8 hrs. of post-feeding (Figure 3 A), whereas in the groups of fish fed on diets FM-M to DC100-AAM supplemented with MPA, the excretion of o-PO4 was lower than those observed in the groups of fish fed on diet FM100 to DC100 But the pattern of excretion of o-PO4 was same as for the groups of fish fed on diets FM100 to DC100. But the pattern of excretion of o-PO4 was same as for the groups of fish fed on the diets FM100 to DC100 (Figure 3 B).
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These results are in agreement with those of Kaushik and Gomes 1998, Kaushik and Cowey, 1991; Van Weerd et al 1993, Jindal 2001, Priyanka and Garg 2002, Kalla and Garg 2004, Jindal and Garg 2005, who reported ammonia and o-PO4 excretion peaks between 7-9 hrs of post feeding.
The differences between the present data and those of others on the peak in ammonia and phosphorus excretion may be attributed to the facilities used by different authors (e.g. the size of holding tanks and the prevailing environmental and rearing conditions). Perhaps it also depends on the species under investigation.
The determination of nutrient budgets and daily patterns of waste excretion in aquaculture is important for evaluating the potential waste load of fish farming effluents (Cowey and Cho 1991). This aspect of intensive fish farming is becoming increasingly important and is receiving much attention (Rosenthal et al 1993).
The decrease in NH4-N and o-PO4 excretion in receiving water with the use of protein of plant origin in feed has important implications on the management of highly intensive farming system. Since the present studies are the initial attempt in this direction, more elaborate work will, therefore, determine the range of applicability.
APHA (American Public Health Association 1998 Standard methods for the examination of water and waste water. APHA, AWWA, EPFC, 20th Edition., New York.
Beveridge M C M and Phillips M J 1993 Environmental impact of tropical inland aquaculture, protein 213-236. In : R S V Pullin, H Rosenthal and J L Maclean (editors). Environmental and aquaculture in developing countries. ICLARM Conference Proceedings 31, 359p.
Bostrom B, Persson G and Broberg B 1988 Bioavailability of different phosphorous forms in fresh water systems. Hydrobiologia 170: 133-155
Buchman J, Sarac H Z, Poppi D and Cowman R T 1997 Effects of enzyme addition to canola meal in prawn diets. Aquaculture 151: 29-35
Cho C Y 1993 Digestibility of feed stuffs as a major factor in aquaculture waste management, protein 365-374. In: S J.Kaushik and P Luquet (editors). Fish Nutrition in practice. Colloques, No. 61. INRA, Paris.
Cowey C B and Cho C Y (editors) 1991 Nutritional Strategies and aquaculture waste. Proc. First Int. Smp. On Nutritional Strategies in Management of Aquaculture waste. University of Gulph, Gulph, Ontario, Canada 1990, p. 275.
Deepak and Garg S K 2003 Effect of replacement of fishmeal with soybean protein on growth and digestibility in the catfish Heteropneustes fossilis (Bloch). Journal of Natcon 15 (1): 131-137.
Garg S K, Kalla A and Bhatnagar A 2002 Evaluation of raw and hydrothermically processed leguminous seeds as supplementary feed for the growth of two Indian Major carp species. Aquaculture Research 33: 151-163
Handy R D and Paxton M G 1993 Nitrogen pollution in Mari culture: toxicity and excretion of nitrogenous compounds by marine fish. Reviews in Fish Biology and Fisheries 3: 205-241
Hastings WH 1976 Nutritional requirement and feed technology. In: Advances in Aquaculture (Editors T V R Pillay, W A Dill) FAO UN Chapter IX, p. 568-609
Jackson A J, Capper B S and Matty A J 1982 Evaluation of some plant proteins in complete diets for the tilapia Sarotherodon mossambicus.. Aquaculture 27: 97-109
Jindal M 2001 Effect of feeding of plant origin proteins on nutrient utilization and growth of Channa punctatus (Bloch.).Ph.D. Thesis. CCS HAU, Hisar pp 141.
Jindal M and Garg S K 2005 Effect of replacement of fishmeal with defatted canola on growth performance and nutrient retention in the fingerlings of channa punctatus (Bloch.). Punjab University Research Journal (Science) Vol. 55, pp 183-189.
Kalla Alok and Garg S K 2004 Use of plant proteins in supplementary diets for sustainable aquaculture. In: National workshop on rational use of water resources for aquaculture. (Editors: S K Garg and K L Jain). March 18-19, Hisar, India, pp 31-47.
Kaushik S J 1992 Recent trends in the development of high-energy diets for salmonids.2e Conference Internazionale “Produione di alimenti per animali”, Piacenza, Italie, 25-26 February 1992.
Kaushik S J and Gomes E F 1988 Effect of frequency of feeding on nitrogen and energy balance under maintenance conditions. Aquaculture 73: 207-216
Kaushik SJ and Cowey CB 1991 Dietary factors affecting nitrogen excretion by fish, p. 7-19. In : Cowey C B and Cho C Y (editors). Nutritional strategies and aquaculture waste. Proc. First Int. Symp. Nutritional Strategies in Management of Aquaculture Waste. University of Guelph, Guelph, Ontario, Canada 1990, 275p.
Kibria G, Nugegoda D, Fairclough R and Lam P 1998 Can nitrogen pollution from aquaculture be reduced? NAGA, ICLARM, 21: 17-25
Kibria G, Nugegoda D, Lam P and Fairclough R 1996 Aspects of phosphorous pollution from aquaculture. NAGA, ICLARM 19: 20-24
Lall S P 1991 Digestibility, metabolism and excretion of dietary phosphorous in fish. P. 163-185. In: C B Cowey and C Y Cho (editors). Nutritional strategies and aquaculture waste. Proceedings of the First International Symposium on Nutritional Strategies in Management of Aquaculture waste. University of Gulph, Ontario, Canada.
Lall S P 1979 The minerals., p. 86-97. In: J E Halver and K Tiews (editors). Fish nutrition and feed technology. Volume I. Heenemann, Berlin.
Pillay T V R 1992 Aquaculture and the environment. Fishing News Books, England. 184p.
Priyanka and Garg S K 2002 Replacement of fishmeal with canola (rapeseed) to enhance growth performance in the catfish Heteropneustes fossilis (Bloch.). Environment and Ecology 20(3): 732-736
Robinson E H and Menghe H Li 2007 Catfish Protein Nutrition (Revised). Bulletin 1153. Office of Agricultural Communications, Mississippi State University, USA
Rosenthal H, Hilge V and Kamstra A 1993 Report Workshop on fish farm effluents and their control in European Community countries, 205p.
Van-Weerd J H, Verastegui A M and Tijssen P A T 1993 Nitrogen excretion and determination of nitrogen and energy budgets in Rainbow trout (Oncorhynchus mykiss, R.) under different feeding regimes. Department of fish culture and fisheries Wagningen Agricultural University, The Netherlands.
Vielman J, Makinen T, Ekholm P and Koskela J 2000 Influence of dietary canola and phytase levels on performance and body composition of large rainbow trout (Oncorhynchus mykiss) and algal availability of phosphorous load. Aquaculture 183: 349-362
Viola S and Lahav E 1993 The protein sparing effect of supplemental lysine in practical carp feeds. Fish-Nutrition-in-Practice. Kaushik S J; Luquet P editors. Paris-France-Institut.-National-de-la-Recherche-Agronomique.
Viola S, Mokady S and Arieli S 1983 Effects of canola processing methods on the growth of carp (Cyprinus carpio). Aquaculture 32 :27-28
Viola S, Mokady S, Rappaport U and Arieli Y 1982 Partial and complete replacement of fishmeal by soybean meal in feeds for intensive culture of carp. Aquaculture 26:223-236
Received 10 July 2008; Accepted 8 September 2008; Published 10 March 2009