Aquacultural Engineering 6 (1987) 183-189

A Tide-generating Apparatus for Laboratory Cultures of Marine Algae

M. Pellegrini and M. Lahaye
Laboratoire de Biologic VEg6tale, Facult6 des Sciences de Luminy Case 901, Universit6 d'Aix-Marseille 2, 70 Route L6on Lacharnp 13288, Marseille C6dex 09, France

ABSTRACT
The apparatus described in this paper may be used for the culture of benthic marine algae or other marine organisms which are submitted to tidal movements. One of the main advantages of our system is that ecological factors (temperature, light, emersion-immersion regimes etc.) can be varied independently from one another and are easily controllable.

INTRODUCTION Marine algae represent an important source of industrial products, especially for their colloidal extracts. Presently, as natural production declines due to overharvesting, it is imperative that intensive seaweed farming should be developed. Therefore, in many countries large seaweed cultivation programs should be seriously considered in order to obtain high quality raw materials for industrial purposes. Different systems of seaweed aquaculture have been attempted successfully either from germlings or from vegetative fragments (see Mathieson, 1975, 1977 for reviews). However, in all cases, for each commercially desirable species, suitable growing conditions must be established, taking into account physical and biological parameters as well as aquacultural engineering problems (for example water supply, pump requirements, aeration, disinfection, etc.). The present paper describes an apparatus which enables researchers to study the effect of varying the ratio of time of emergence/time of immersion on the growth of various seaweeds cultivated in a free-living state.
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DESCRIPTION OF THE APPARATUS The culture apparatus consists of 12 modules supported by a metallic frame, arranged in two thermostatically controlled water units which are equipped with fluorescent temporized lighting (Fig. 1 ). The complete change of culture medium is carried out automatically by simple gravity through a distribution tank. A schematic diagram of the whole system is represented in Fig. 2; its components are described separately below. Each unit is composed of three main elements: (a) an upper culture tank, (b) a central transfer tank and (c) a pump located in the lower part.

Fig. !.

Photographic view of the tide-simulating apparatus. (a) culture tank; (b) transfer tank; (c) lower pump.

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"

Fig. 2. Schematic view of the tide-simulating apparatus. (a) aerator; (b) basket; (ct) culture tank: (dp) distribution pump; (dt) distribution tank; (fit) fluorescent light tubes: (LLW) lower level of water; (lp) lower pump; (lwp) lower waste pipe; (op) overflow pipe: (p) pump; (r) reservoir; (s) siphon; (tb) thermostated baths; (tt) transfer tank; (ULW) upper level of water: (UV) ultraviolet sterilization system.

Seawater enters the culture tank (20 litres) through a plastic tube connected to the lower pump. An overflow pipe marks off the maximal level of water. A siphon allows partial drainage and induces the +low-tide" condition. Constant motion of the culture medium is provided by bubbling air from the bottom of the tank through a pump (Rena, model 325, flow rate: 6.5 litres min- ~) and an aerator. The specimens are placed into four plexiglass baskets which are perforated on their sides and suspended by adjustable clamps from the top of the culture tank.

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The central transfer tank (20 litres) is connected to the overflow pipe and the siphon of the culture tank. It is provided with: (a) a delivery water pipe linked with the distribution tank, (b) an overflow connected to the laboratory waste pipe and (c) a siphon which maintains the lower pump under load. The distribution pump (Eheim, model 2013, flow rate: 390 litres h-J) of the culture tank, fitted with an active charcoal filter, is equipped with a timer. A reservoir (450 litres) contains filtered seawater which passes through an ultraviolet sterilization system, in order to minimize microbial contamination, before reaching the transfer tank by gravity. This ensures that the tide rates are not disturbed in the culture tank. The sterilization system consists of a quartz glass tube discharging 2 537 A ultraviolet radiation. The six culture and transfer tanks are placed in two water baths which are maintained at the same temperature. Temperature regulation is provided by a thermostat located in the bath and connected to a freezing apparatus and to a pump which circulates ethyleneglycol through coils at the bottom of the thermostated baths. All tanks, baths and pipes are made from plastic or polyvinyl chloride (PVC) which is unaffected by seawater, in order to avoid metallic ion contamination. Moreover, it is known that plastics do not inhibit algal growth (Dyer and Richardson, 1962). Illumination is provided for each water bath by four 65W fluorescent tubes (2 Gro-lux Sylvania alternated with 2 cool-white) mounted on a board and suspended above the front end of the culture system. The energy emission of these combined fluorescent tubes closely follows the absorption spectrum of algal pigments. The light intensity may be modified by changing the tube number. It is possible that some evaporation of the medium takes place, leading to an increase in salinity. It is therefore imperative to monitor salinity daily to maintain it at a constant level. The medium is changed regularly each week. Culture units are installed in a constant temperature room.

OPERATION Tidal conditions are maintained automatically in the following manner. The lower pump of the unit draws water from the transfer tank and supplies the culture tank causing the water level to rise, simulating 'flowing tide'. The water reaches the overflow; this corresponds to the

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'slack high water'. When the pump stops, the siphon returns the water to the transfer tank, simulating 'falling tide'. When the siphon fails, it is 'slack low water'. Then the cycle starts again.

DISCUSSION Cultures of free-living algae are generally maintained in motion by producing water circulation with an air-jet sunk into the culture tank. This technique has been used for different Rhodophycophytes, especially for Chondrus (Neish and Fox, 1971; Neish and Shacklock, 1971; Neish et al., 1977) and for Gracilaria (De Boer and Ryther, 1977). It is known as 'tumble culture' and differs from the 'spray culture' technique developed for algae attached on frameworks, suspended vertically and immersed in a spray of sea water (Chapman, 1973). Intertidal algae undergo different periods of immersion and, according to Doty and Archer (1950), the tide must be considered to be a primary factor regulating secondary factors such as temperature, light, etc. However, inside an experimental culture unit, the motions of the water caused by tides are difficult to reproduce as are those caused by waves and currents. South (1970) experimented with a free-flowing sea water system for the culture of Alaria, whereas Charters and Neushul (1979) built an apparatus using a submerged jet of high velocity water which vigorously brushed attached algae for a short time period. Three principal types of laboratory apparatus have been constructed for simulating tidal action (see Underwood, 1972, for a critical review): ( 1 ) Hydrographical models which allow the study of controlled physical factors such as wave form and tidal velocity. These are rarely used by biologists (Anon, 1967, 1968; Thom, 1960). (2) Siphonal models which produce alternate raising and lowering of water in culture tanks by means of a system of drains and valves (Bracher, 1919; Martin and Reed, 1935; Aleem, 1949; Barkman, 1955; Fulcher and McCully, 1969; Rusanowski and Vadas, 1973; Fletcher and Jones, 1975). Unfortunately such apparatus suffer from some disadvantages, for example, the change in water level occurs at a constant rate whereas it follows a sine curve on the seashore. (3) Sinusoidal models which accurately simulate the natural conditions by producing sinusoidal changes in water level (Evans, 1964; Micallef, 1967; Thompson, 1968; Townsend and Lawson, 1972: Underwood, 1972; Edwards, 1977).

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Our system, easy to build and operate, is based on the 'tumble culture" technique in that an attempt is made to artificially reproduce tidal action in laboratory studies. Culture experiments are currently being conducted to investigate the effect of time of emergence relative to time of immersion under varying regimes upon the growth of fragments of a brown alga Cystoseira stricta, the regenerative properties being used as a means of increasing the natural yields of the plant. This Fucale is distributed on the exposed rocky shores of Provence, in dense bands, just below the water fiaark so that it is covered and uncovered due to wave and tidal movements, even though these are very feeble in the Mediterranean Sea. The effects of nutrient addition are also being investigated. Complete results will be published elsewhere. In conclusion, it is obvious that the main advantage of our experimental apparatus is that it is easily controllable. Indeed a variety of combinations of temperature, light intensity, photoperiod, emergenceimmersion regimes may be investigated in order to define optimum cultivation conditions for benthic marine algae or other marine organisms.

ACKNOWLEDGEMENTS The authors wish to thank G. Slawyk (Centre d'Oc6anologie de Marseille) for criticizing the text and for assistance in the preparation of the English text.

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