Aquacultural Engineering 4 (1985) 235-246

A Laboratory Scale Recycling Water Unit for Tilapia Breeding

M. Koiller a n d R . R . A v t a l i o n Department of Life Sciences, Bar-Ilan University, Ramat-Gan 52 100, Israel

A BS TRA CT The technical features o f a laboratory scale water recycling unit for experimental small scale tilapia breeding are described. Two units (1 and 2) were operated during a 6 month period, carrying a similar fish load (7.5 kg) and feeding rate (2% fish body weight~clay). Unit 1 received natural illumination, while unit 2 was artificially illuminated (14/10 - light/dark cycle). Both units were equipped with a biological filter bed (substrate surface area, 3500 era2). In unit 1, total ammonium and nitrite concentrations ranged from 0.05 to 0.5 mg liter-a, while nitrate varied between lO-40mg liter-~. In unit 2 corresponding values were 0.15-3 mg liter-1, 0.05-0.8 mg liter-~ and 10-40 rag liter-~. Temperatures ranged between 20-29C and pH values between 7.5-6.9 in both units. Dissolved oxygen concentrations decreased gradually from 5.6 to 3.4 mg liter-~ in unit 1 and from 5.6 to 2.6 mg liter-1 in unit 2. Twenty-six spawnings occurred in unit 1 in March and April, while only eight spawnings occurred in unit 2, possibly because o f the absence o f sunlight. The significance o f these results are discussed.

INTRODUCTION The continuous elimination of toxic metabolites and growth-inhibiting substances is an essential process when operating a closed system for intensive fish culture. Many recycling water systems using biological filters have been used for the degradation of accumulative organic compounds which originate from fish excretion and excessive feed (reviewed by Otte and Rosenthal, 1979). However, biological filtration has some disadvantages, in particular, the accumulation o f nitrate (the end-product of nitrification) and low-biodegradable substances. To 235 Aquaeultural Engineering 0144-8609/85/$03.30 - Elsevier Applied Science Publishers Ltd, England, 1985. Printed in Great Britain

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cope with this problem a certain amount of water must be continuously replaced or the recirculated water treated accordingly (i.e. anaerobic denitrification, activated charcoal, ozonation). A further disadvantage of biological filters is their slow adaptability to environmental variations, such as changes in temperature, salinity, dissolved oxygen, organic waste load and fish biomass (Coldberg and Lingg, 1978; Otte and Rosenthal, 1979). Nevertheless, when operating small closed systems for fish breeding on a laboratory scale, it seems reasonable to employ biological filters and to change the water partially, whenever necessary, rather than to treat it chemically (chemical filtration devices, ozonation, etc.). The accumulation of organic residues and the harmful effects of these residues on fish growth is one of the main problems of intensive aquaculture. The sedimentation of organic matter in fish ponds results in anaerobic conditions and the production of toxic materials such as NH4, H2S, etc. The continuous aeration and resuspension of organic matter was reported to enhance the recycling of the organic matter in fish ponds (Avnimelech and Lacher, 1980; Shilo and Rimon, 1982). Similar processes occur in biological gravel beds. The accumulation of heavy detritus (particulate matter) in the gravel bed increases the biological oxygen demand (BOD) of the system and reduces its carrying capacity. Hence, excessive detritus must be removed periodically (Spotte, 1979). In this article we describe a laboratory scale recycling unit for experimental breeding of tilapias. This unit is equipped with a biological gravel-bed filter capable of avoiding excessive accumulation of detritus and thus avoids supporting effective nitrification.

MATERIALS AND METHODS The recirculation unit consisted of four aquaria and an external biofilter (see Fig. 1). Aquaria of 250 liters (169 X 38 X 39 cm) provided space for a family of tilapia composed of 2 males and 4-6 females. (Each aquarium had 7-8 fish with an individual average weight of 250g, totalling 1875 g of fish. The average individual fish length was 25 cm.) Two separate nesting zones were established by the 2 males in the corners of the aquaria. Aquaria consisted of a skeleton of fiberglass to which the walls

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and the bottom plate were glued. The front wall was made of glass (8 mm thick), while the bottom and the rest of the walls were made of asbestos (8 mm thick). The level of the water (33 cm) in each aquarium was maintained by an overflow outlet, from which the collected excess water was piped into the biofilter bottom. The biofilter (70 50 40 cm) and its components are depicted in Fig. 2. Waste water from aquaria flows through a PVC collecting pipe (5 cm diameter) into the sediment settling space (see Fig. 2) where the first stage of filtration occurs. Since the filtration process proceeds from the bottom to the top (upflow speed 1-14 cm min-1), heavier particulate matter precipitates when the water in the sediment settling space flows upwards through the gravel bed where the biological filtration takes place. The filtered water is recirculated to aquaria by a submerged centrifugal pump (Eheim Filter & Fish Co., Germany). The level of the water in the biofilter is maintained by a float valve. When a complete or partially open system is operated, the excess water flows through the overflow to the sewage. The first stage of filtration represents a significant reduction step of the organic load, since accumulated suspended solids are evacuated to the sewage at desired intervals using a programmed automatic tap (solenoid valve). A mechanical tap was

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fixed before the solenoid valve to permit manual evacuation. This system, besides permitting the replacement o f water at the desired rates, also allows self-cleaning o f the filter and avoids its overloading since, during the evacuation time the filter bed is subjected to a reverse-flow. The gravel bed and components are illustrated in Fig. 3. The gravel bed is composed o f two layers of gravel separated by nets. The first layer, 5 cm thick, consists o f dolomite gravel o f 2-5 mm grain size, with a void space o f 360 ml liter -1 -+ 0.83 and an inner surface o f 5760 m 2 m - 3 + 2-32. (To simplify the inner surface calculation, the gravel was assumed to be spherical.) The second layer 4 cm thick, consists of a porous gravel (Effisubstrate - E h e i m Filter & Fish Co., Germany) of 5 mm grain size, with a void space o f 538.6 ml liter -1 -+ 0.53, thus offering an ideal and extended adhesion surface for bacteria. This layer is completely isolated from the second one, since it is enveloped by a white nylon net o f 0.8 mm mesh size that also mechanically filtrates the water. However, the inner surface of this porous gravel, u n d o u b t e d l y

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much more extended than the dolomite gravel, is difficult to calculate. A 3-mm mesh net separates the dolomite gravel from the white net. The dolomite gravel [CaMg(CO3)]2 acts as a natural buffer in the water system since it contains a large source o f carbonates to neutralize the acid-forming processes o f closed culturing systems (Spotte, 1979). The gravel bed with the filtration elements can easily be dismounted. Recirculation o f water is described in Figs 1 and 2. A tap (T) was connected to a pipe (1-27 cm diameter) to allow free flow of the recirculated water, avoiding head loss o f the pump. Four taps were connected to this pipe to supply the recirculated water to each aquarium. A tap was connected to the system to provide water from an external source. If a partially open system is desired (e.g. amounts o f water equivalent to 20-30% day -1 of the total recirculated water in the closed system), this tap is regulated to supply water. It is also possible to operate a completely open system for some hours by opening the external water source tap. In that cae, the tap (T) must be closed to prevent water flowing towards the biofilter. In addition, the biofilter sewage tap and the solenoid valve must be opened. In that way, the waste water entering the biofilter, instead of flowing upwards through the gravel bed and filter overflow, would flow directly from the sediment settling space to the drain system. The centrifugal pump was regulated to circulate the water at a rate of 4 liter min -1 (1 liter min -~ inflow rate to each aquarium). Since the void space of the biofilter gravel bed equals 13-84 liters, the retention time of the water in the biofilter can be evaluated to 3.5 min. Recirculation occurred every 4 h, allowing a theoretical turnover of 6 times day -~. Aeration to aquaria was supplied b y air diffusers and airstones (see Fig. 4). The diffuser (Eheim Filter & Fish Co., Germany) was fitted at the end o f the inlet hose on the water's surface and attached to the aquarium wall (spray angle a b o u t 45). The air-hose of the diffuser is taken upwards to a point outside the aquarium. The diffuser unit mixes the supplied air with the incoming water, which is sprayed as a result of the pressure of the water. This ensures oxygenation o f the water and enables surface agitation, an important factor for diffusion of oxygen across the air-water interface. The diffuser provides a good indication of the flow rate of the system. When the pump or the taps are blocked with particles of matter, the reduced water pressure reduces the quantity o f air sprayed by the diffuser. In case o f pump failure, the diffuser would not work. Simultaneously, a continuous aeration to

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aquaria is supplied b y a compressor through a separate pipe (see Fig. 1) connected to an airtap and airstone. The water temperature (26-28C) is maintained b y thermostatically controlled immersion heaters. The feeding rate was established as 2% o f the average b o d y weight of the fishes day -x , but during experimentation an attempt was made to increase this percentage. The fish were fed with commercial trout pellets containing 42% proteins, 12% lipids, 20% wheat flour, 1% vitamins and 2% cellulose. The water quality measurements were taken without the operation of the solenoid valve at a 1 liter min -1 flow rate to each aquarium and the mechanical tap was opened once a day to replace about 10% of water content o f the biofilter container. Measurements of temperature, pH, dissolved oxygen, ammonia, nitrite and nitrate were tested. These determinations were carried out using the m e t h o d o l o g y described in the literature (Taras et al., 1971 ; Boyd, 1979).

RESULTS Water samples for quality determinations were initially taken in two independent units (units 1 and 2), once per week, usually before feed-

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ing (during morning hours). Unit 1 was installed in a place where the roof was partially transparent and near large windows allowing strong natural illumination and heat (essential factors for the spawning process in tilapias), Unit 2 was artificially illuminated by fluorescent lighting ( 1 4 / 1 0 - light/dark cyele). Both units began operating in December, 1982. The determinations are shown in Figs 5 and 6. Temperatures in units 1 and 2 varied between 20-29C and 20-28C, respectively. These variations were more pronounced in winter because of the sharp shifts in environmental temperature and the insufficiency of the heaters used to provide the desired temperatures during the colder days. Dissolved oxygen levels of the filtered water decreased from 5-6 to 3.4 mg liter-1 in unit 1, and 5.6 to 2.6 mg liter-1 in unit 2, probably due to the oxygen consumption of microorganisms decomposing organic matter in the gravel bed. In addition, heavy detritus covering the gravel bed (especially noted in unit 2) and accumulative organic load in the culture system, offered additional substrate for bacterial growth and consumption of dissolved oxygen. Furthermore, the level of dissolved oxygen decreased when the pump or the taps and diffusers were partially blocked by particulate matter. The pH values decreased gradually with time, varying between 7.56.9. This decrease is the result of alkalinity-loss during nitrification and oxidation of organic waste products by bacteria. However, the partial water replacements neutralized such decreases. It is possible that the dolomite gravel contributed to the gradual decline of the pH. Total ammonium concentrations in unit 1 ranged between 0.05-0-5 mg liter-x and heavy detritus covering the gravel bed was not observed as in unit 2. In unit 2, ammonium values were kept between 0.15-1 mg liter -1, but at the end of the experiment a sudden increase (up to 3 mg liter-~) was observed. Even though water was changed, total ammonium levels continued to rise. This unusual increase of ammonium can possibly be explained by a significant reduction of the oxidative capacity of nitrifying bacteria in the gravel bed, due to the interspecific competition with heterotrophic bacteria. For both units, the low pH values assured that unionized ammonia was present at very low levels, being harmless to the cultured fish. Nitrite and nitrate values in unit 2 varied from 0-05 to 0.8 mg liter-~ and 10 to 40 mg liter-x, respectively, while in unit 1 they were 0.05 to 0.5 mg liter-1, and 10 to 40 mg liter-1, respectively.

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DISCUSSION The concentration of total ammonium ions increased with time, accompanied by nitrite and nitrate increases in both units, indicating some incomplete nitrification. In unit 2, ammonium and nitrite reached maximum levels (1 mg liter-1 and 0.8 mg liter-1, respectively) after almost 9 weeks without water replacements. This can largely be attributed to the fact that the biofilter was covered with excessive detritus supporting heterotrophic bacteria. The aquarium water presented a very strong yellow color, indicating the accumulation of yellow substances and other low-degradable organisms. The activity of fish was reduced and one case of death occurred. In unit 1, however, both ammonium and nitrite values only reached 0.5 mg liter -1. The filter was slightly covered by detritus, the fish activity was normal and no mortality was experienced. It is possible that the low-degradable substances in connection with lowered oxygen (unit 2) may have affected the fish rather than nitrite alone at the concentration of 0-8 mg liter -a. It is also possible that nitrite toxicity was enhanced as a result of the combined effects with the other accumulated metabolites. In both systems the increases in temperature generally coincided with the increase of metabolite. This shows, that metabolic excretion and activity increased faster with temperature rise than the biofilter activity. Fish breeding activity was reduced at low temperatures and, consequently, the organic load and the metabolite levels were reduced. At 23C fish could consume more, but at 25C the feeding activity was greatly intensified. When the temperature varied between 25-29C, the feeding rate was increased to 3% fish bodyweight/day-1, but some feed remained at the bottom of the aquaria. Therefore, the increasing temperature resulted in a higher fish metabolism for which the bacterial activity in the filter was not sufficient. Spawnings were more intense in unit 1 due to the natural illumination and the higher temperature regime (26 spawnings during March and April), while only eight spawnings occurred in unit 2. The partial water replacements accelerated the spawning process and handled the metabolites at lower levels. The results of the water quality determinations for both units show that the biological filter could not handle the accumulation of metabolites and low-biodegradable substances (yellow color in aquaria) without water replacements, although the total bioload was relatively

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low. A water replacement rate o f about 50% of the total volume o f the unit, every 3 to 4 weeks, was found to be satisfactory. In order to maintain the same water quality conditions that prevailed during this experiment and to always guarantee sufficient total ammonia oxidation for the total biomass load over longer time periods, in a permanent closed system, there is a need to determine the carrying capacity o f the biofilter. Hirayama (1974) proposed a formula for the calculation o f the carrying capacity o f a small marine system. However, this formula could not be applied to our system because we still cannot calculate the inner surface o f a porous gravel, which may enlarge the surface filtration area for bacterial attachment. Hirayama stated that in his method of estimating carrying capacity, the water purification was assumed to occur only on the surface o f the sand grains. A higher flow rate (higher than 1 liter min -1 per aquarium) can reduce the BOD load (Otte and Rosenthal, 1979) as can maintaining the feeding rate at only 2% day -1. This will possibly improve the performance of the biofilter.

ACKNOWLEDGEMENTS This work has been supported b y grant 03.509 from The Research and Development Co. Ltd, Bar-Ilan University, and b y grant AQ 24 from the GKSS, Gessthecht-Tesperhude, FRG. We are sincerely grateful to Dr Harald Rosenthal, o f the Biologische Anstalt Helgoland, H a m b u r g Bahrenfeld, F R G , for his advice and help in editing this manuscript. REFERENCES Avnimelech, Y. & Lacher, M. (1980). On the role of soil in the maintenance of fish pond's fertility in development of hydrobiology. In: Development in Hydrobiology, vol. 2, eds J. Barica and L. R. Mur, Junk Publ., Holland. Boyd, C. E. (1979). Water quality in warm water fish ponds. Auburn University Experimental Station, Dept of Fisheries and Allied Aquaculture, Auburn University, Alabama, pp. 201-76. Coldberg, P. J. & Lingg, A. J. (1978). Effect of ozonation on microbial fish pathogens, ammonia, nitrate, nitrite, and BOD in simulated reuse hatchery water. J. Fish. Res. Bd. Canada, 35, 1290-6. Hirayama, K. (1974). Water control by filtration in closed culture systems. Aquaculture, 4,369-85.

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Otte, G. & Rosenthal, H. (1979). Management of a closed brackish water system for high density fish culture by biological and chemical water treatment. Aquaculture, 18, 169-81. Shilo, M. & Rimon, A. (1982). Factors which affect the intensification of fish breeding in Israel. 2. Ammonia transformation in intensive fish ponds. Bamidgeh, 34, 101-4. Spotte, S. (1979). Fish and Invertebrate Culture - Water Management in Closed Systems, John Wiley & Sons, New York. Taras, M. J., Greenberg, A. E., Hoak, R. D. & Rand, M. C. (Eds.) (1971). Standard Methods for the Examination o f Water and Waste Water, APHA, AWWA and WPCF Press, 13th edn, pp. 232-48.