aquacultural engineering

ELSEVIER Aquacultural Engineering 16 (lYY7) 161-166

The influence of feed protein intake on tilapia ammonia production

J.L. Brunty, R.A. Bucklin, J. Davi$, C.D. Baird, R.A. Nordstedt
Agricultural und Biologicul Enginruring Drpartmmt, Uniwrsi~ of Florida. Gainrs~illr. FL 3261 I. USA 72hnology Group. EPCOT Center: Luke Bwna Vista. FL 32830. USA Received 5 September 1996; accepted 8 November 1996

Abstract
Biological filters designed to remove toxic ammonia excreted as fish wastes are an integral part of any recirculating aquacultural system. The quantity of ammonia excreted by fish is related to the quantity of nitrogen supplied by the protein contained in feed. This experiment evaluated short term Total Ammonia Nitrogen (TAN) production by juvenile tilapia fed rations with six different protein contents. Five feeds composed using soybean meal as their protein source and one feed deriving most of its protein content from fish meal were tested. Statistically significant increases (P~0.10) in TAN production were observed as protein content increased during the 24 h period following feeding. Fish meal and soybean meal did not produce statistically different TAN production rates. 0 lYY7 Elsevier Science B.V. ki~~orrls: Feed protein; Tilapia: Ammonia

1. Introduction
Aquaculture is becoming an increasingly important source of fish products as harvests from natural waters decline worldwide due to overfishing and pollution. Currently, most fish are grown in outdoor pond systems where water is diverted from a nearby lake or river for a single pass through the pond system and then released again to the environment. The quality and the quantity of water used are becoming an increased concern and efforts are being made to develop production

0 144~X609/97/$17.00 0 1997 Elsevier Science B.V. All rights reserved.

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systems wherein the water is at least partly recycled. The quantity of nutrients added by fish wastes is a major concern when designing systems to reuse water. Total ammonia nitrogen (TAN) levels can be a limiting factor for the reuse of water because high levels can reduce fish vitality and/or growth or cause death in extreme cases. Protein in fish food is the primary source of nitrogen that is excreted through fish gills as ammonium ions. Knowledge of protein utilization rates will allow selection of the most economical feed mix to minimize protein wastes and reduce both costs and ammonia production. This study attempted to evaluate the relative levels of TAN production that resulted from varied sources and levels of feed protein.

2. Background Protein within the fish feed is the source of ammonia introduced into production systems and higher levels of the nitrogen source should result in higher levels of TAN production. However, different species of fish and fish at different life stages within the same species are differentially able to utilize the protein in feed, and the relationship between protein levels and TAN production for all species and all life stages may vary (Begum et al., 1994). Proteins are generally considered to contain 16% N (Overton, 1993). The reason for quantifying protein requirements is to develop the lowest cost feed that provides adequate or maximum growth. Animal sources of protein are typically more expensive than plant sources (Wee and Shu, 1989). Fish meal is typically composed of 25-65% protein, while plant sources only contain 5-15% protein, therefore necessitating the use of fish meal as a protein base. Speece (1973) observed that from 0.026 to 0.032 lb NH3-N was produced per lb of feed at temperatures from 49 to 62F (26 to 32 g of NH3-N per kg of feed at 9.4 to 16.7C). Several studies (Khan et al., 1993; Tucker and Boyd, 1979; Santiago and Reyes, 1991; Watanabe et al., 1993; Reigh and Ellis, 1992; Wu et al., 1994 have investigated the effects of feed source and protein content on the quantity of ammonia eventually released by the fish into production water, but did not report ammonia levels in terms of feed protein content. Studies relating ammonia levels to feed protein content are not in close agreement. Masser et al., 1992; Meade (1985); Mires and Amit (1990); Rychly (1980) and Wright (1993) report that fish excrete from 60 to 95% of feed nitrogen as ammonia and that 1 kg of feed will produce 19.5 g of NHX-N and possibly as much as 300 g NH4-N.

3. Materials

and methods

This study was conducted in the Structures and Environment Lab of the Agricultural and Biological Engineering Department at the University of Florida over a 4 week period during the fall of 1994. Two groups of young hybrid tilapia were

.I. L. Brmty rt al.lAcpacultural Enginerring 16 (1997) 161-166 Table I Feed composition Feed 32%) protein Arkat Minnow Meal Factor Crude protein Crude fat Crude fiber Protein source > 32?+> ;3.5% 5 0% Soybean meal 2 34%) 2 696 Soybean meal z 36% 235% 55% Soybean meal > - 38% 2 10% Soybean meal > 42% 2 7% 239; Soybean meal 34% protein Ziegler Cichlid Diet 36% protein Arkat Minnow Meal 38% protein Ziegler Tropical Fish Feed 42% protein Arkat Minnow Meal 55%

163

protein Ziegler Salmon Starter

> 5S%, 2 15% < _ 2% Soybean meal

placed in two separate 37.9 L tanks, with 21 fish (160 g total weight) in Tank 1 and 20 fish (99 g total weight) in Tank 2. Fish were fed the feed type to be tested for 24 to 48 h prior to the beginning of each feed test. The composition of the feeds tested, as described on the manufacturer s label for each feed, are listed in Table 1. Each group was placed in a tank of fresh tapwater dechlorinated through the use of NovAqua Tapwater Conditioner and fed 2% of the overall average body weight to begin the test. The fish were then left in a lighted room overnight. Ammonia measurements were taken at four separate times the next day. At the end of the day, the fish were placed in another clean tank and again fed 2% of the overall body weight of the same feed and the process was repeated. The fish were fed the next feed type to be tested for l-2 days before the next feed test began. Ammonia levels were measured using Nessler s Reagent and Hach Company Nessler Method (Hach, 1993) on a Milton Roy spectrometer. The method measured total ammonia nitrogen (TAN) including both NH3 and NH,+. The relationship between ammonia production levels and the nitrogen contents of the different fish feeds was statistically analyzed using the Regression Procedure of the SAS System (Statistical Analysis Systems Institute Inc., 1990). An analysis of variance was conducted using the General Linear Models Procedure of SAS to evaluate time effects and differences in dissolved oxygen, water temperature and water pH. Differences at the 90% probability level or greater were considered to be statistically significant. 4. Results Water temperatures ranged between 24.0 and 25.2C, with fluctuations occurring at approximately the same time in both tanks. Dissolved oxygen levels throughout

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J. L. Brunty et al.lAquacultural

Engineering 16 (1997) 161-166

the study were above 5 ppm in both tanks with the exception of Day 1 of the 32% protein feed in Tank 2 when dissolved oxygen measured 4.3 ppm. The fish grew from initial weights of 7.6 g per fish in Tank 1 and 4.9 g per fish in Tank 2 to final weights of 11.1 g per fish in Tank 1 and 14.7 g per fish in Tank 2. No fish died during the study, and the final total weights were 233 g in Tank 1 and 294 g in Tank 2. No statistically significant differences related to treatments were observed in pH. The average pH was 7.04 with a standard deviation of 0.12. Results of the regression of observed TAN versus nitrogen content of feed indicated that ammonia levels increased with significant (P ~0.10) linear trends as protein levels increased. The linear increase in TAN production as a function feed nitrogen content (FEED-N) is expressed by Eq. 1 where TAN and FEED-N are expressed in grams per kilogram of feed. The coefficient of determination (?) for this relationship is 0.68. TAN = 0_604*FEED-N+3&3 (1)

The experimental observations for all feed trials and the values predicted by eqn (1). are shown in Fig. 1. Statistical analysis determined that there was significant

MEAN TAN +/- ONE STANDARD DEVIATION

TAN = 0.604*FEED-N + 3.88

FEED-N, grams per kilogram

Fig. 1. Observed TAN levels.

of feed

J. L. Brunty et al.lAquaculturul Engineering 16 (1997) 161-166

165

Table 2
Observed TAN levels

Feed protein content Grams of nitrogen per kilogram of feed Grams of TAN measured per kg of feed (SD)

32c7r Sl.1 35.1 (1.1)

34% 54.4 40.x (4.1)

36% 58.7 34.1 (2.1)

38% h@X 39.7 (4.2)

424 67.2 44.9

5SCi 884 57.4

(24)

(?4)

variability in TAN levels as a result of the various feed treatments. Table 2 lists the feed N contents and the observed TAN levels.

5. Discussion

Ammonia levels increased linearly as protein content increased, without regard to protein source. The coefficient of determination of 0.68 indicates that a linear model can explain 68% of the relationship between TAN and feed protein content. Ammonia levels were expected to increase with feed protein content because the nitrogen content of feed increases as the feed protein content increases. Increased nitrogen content results in increased potential for ammonia formation because of the increased presence of the nitrogen atoms required to form ammonia. Increasing ammonia levels as a result of increased feed protein content agrees with the findings of Wright (1993) Meade (1985), and Rychly (1980). Feed protein source had no significant effect on ammonia production in this study. Ammonia production from fish meal proteins were within the limits of the linear model developed by the statistical analysis. The results may differ for long term feeding effects between fish meal and soybean meal proteins. Brunty (1995) found that over an 8-week period, the 40% Silver Cup and 42% Arkat feeds used in this experiment resulted in significantly different TAN, pH, and nitrate levels. Because the protein levels in Brunty (1995) were nearly equivalent, these differences were attributed to the source of the protein content. Because the fish were fed a minimal percent of their body weight (20/o), all of the feed was consumed. The average percentage of nitrogen excreted for all feed types was 66% compared with the value of 80% found by Mires and Amit (1990). Nitrogen excretion levels observed in this study ranged from 34.2 to 57.4 g of TAN per kg of feed compared to the values of 19.5 g of NH3-N per kg of feed reported by Meade ( 1985) and 26 to 32 g of NH3-N per kg of feed reported by Speece (1973). Many factors that may affect TAN production, such as temperature, salinity. alkalinity, fish species, dissolved oxygen, etc. were not considered in this experiment. These factors alone or in combination with other factors may cause a different protein content/TAN level relationship when the factors are different from those found in this study.

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