Aquacultural Engineering 2 (1983) 279-286

An Apparatus for Long Term Biological Experiments in Running Water at Temperatures Close to Freezing G. Sivertsen, J.C. Wallace
Institute of Fisheries, University of Troms, PO Box 3083, Guleng, N-9001 Troms, Norway

and
T.G. Heggberget Directorate for Wildlife and Freshwater Fish, Sverresgt. I, N-7000 Yrondheim, Norway

ABSTRA CT An open aquarium system in which water temperatures can be manipulated and controlled close to freezing point has been constructed. Technical details and the results o f a working trial lasting six weeks are reported. The equipment was constructed in order to be able to simulate river water temperatures, and changes in these due to hydroelectrical development in Arctic regions. The incubation o f Atlantic salmon eggs was o f particular interest. Other applications include growth studies and nutritional physiology o f aquatic organisms at low temperatures. The system can be used with both freshwater and seawater.

INTRODUCTION In order to study experimentally the relationship between water temperature and growth or development of aquatic organisms it is necessary to be able to control water temperature in an experimental system accurately. If the system is an open one, i.e. through flow rather than circulation, and the required temperatures close to zero, this becomes technically difficult, since any unwanted ice formation may make temperature control thereafter impossible. This may explain the fact that most experimental data on the aforementioned relationship 279 Aquacultural Engineering 0144-8609/83/$03.00 Applied Science Publishers Ltd, England, 1983. Printed in Great Britain

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G. Sivertsen, J. C. Wallace, T. G. Heggberget

has been obtained from experiments carried out at temperatures no lower than 2C. However, in northern temperate and Arctic regions running waters are in winter relatively shallow water layers which run under ice, and they are in consequence cooled to temperatures close to freezing for periods of up to several months. It is under such conditions that river organisms live during a significant part of their life cycles, and it is, therefore, important to be able to undertake long term physiological experiments at these low temperatures in order to gain insight into the biology of these species. This paper describes an open system in which the water temperature in relatively large holding tanks or aquaria can be accurately controlled to as low as 0.1C or lower (if salt water is used). DESCRIPTION OF A P P A R A T U S (See Appendix for Equipment Specification) Figure 1 shows a diagram o f the apparatus. Water is taken from a header tank placed at a height of about 2 m above the experimental tank. This ensures the even flow which is essential for temperature control. F r o m the header tank the water is gravity fed through a 20 mm PVC pipe to an insulated pre-cooling tank placed above the experimental tank. The pre-cooling tank is an oval plastic tank placed inside a larger one, the space between them being packed with insulating material. With a surface-to-surface water level difference of 1.5 m the water overflows from the pre-cooling tank to the experimental tank. This is an Ewos 1 m 2 by 45 cm deep glass fibre tank with a telescopic depth c o n t r o l / o u t f l o w and a central b o t t o m drain. It is insulated on the sides and base with 5 cm thick expanded polystyrene. Water flowing into the experimental tank is distributed around the sides through small jets, 2 mm holes drilled on the underside o f a PVC pipe m o u n t e d just above the cooling coil. At low flow rates it is conceivable that the flow within the experimental tank could be uneven. To prevent this the central drain has been covered by a plastic plate, with a very small clearance. The slightly sloping tank b o t t o m has also been made completely horizontal b y the addition o f a mat o f plastic foam. On top o f this mat is a layer o f sand

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Fig. 1.

COOLING COIL Principal design of the cooling system. (For technical details see text.)

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G. Sivertsen, 3". C. Wallace, T. G. Heggberget

(particle size 1-3 mm) 3 cm thick, and on top o f the sand a perforated aluminium plate, which is n o w the effective tank b o t t o m .

Cooling system
The water in both the pre-cooling tank and the experimental tank is cooled by a glycol solution, diluted to give it a freezing point o f - - 2 2 C . It is circulated by pumping from a reservoir through separate cooling coils o f 25 mm PEH pipe in the two tanks. In the pre-cooling tank the cooling coil lines the walls with a clearance o f 2-3 cm. In the experimental tank the coil is placed on the water surface. This coil has 10 windings and the clearance between the tank walls and the outermost winding is 5 cm. The coil is held in place by an aluminium clamp corlstructed as a cross with spacers to hold the windings apart. The cross is suspended by ropes, and the coil is coupled to the glycol system by flexible hoses.

Temperature control
To obtain accurate temperature control in the pre-cooling tank the glycol cooling coil is made to operate against a warming unit. This is a 'Hetotherm' temperature control unit (effect 1200/600W) which stirs the water vigorously and which controls the temperature to an accuracy of 0 . 0 1 - 0 . IC by means o f an adjustable mercury contact thermometer. The temperature o f the glycol solution can be controlled in a number o f ways, viz. (1) by the cooling compressor's thermostat; (2) by control either manually or automatically, of the circulation pump; (3) by regulating the amount of glycol flowing into the coils by means of flow control valves.

Operation
At experimental temperatures higher than IC the cooling coil in the experimental tank is not used. A temperature sensor on the b o t t o m of the tank monitors b o t t o m temperature and the water temperature in the pre-coohng tank is simply adjusted to give the desired temperature in the experimental tank. (This usually involves adjusting the precooling tank temperature slightly lower than that required in the

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experimental tank, since even with well insulated tanks and water hoses there is usually a slight warming as the water flows from the first tank to the second.) When water temperatures lower than 1C are required in the experimental tank, glycol is circulated through the horizontal cooling coil. This results in the formation of an ice plate on the surface of the tank. A timer regulates the length of the period during which glycol is pumped through the coil and, therefore, the thickness of the plate. In addition, the inflow of water all around the tank prevents the ice plate from extending to contact the walls and so disturb the water flow pattern. The depth of water in the experimental tank, and therefore the distance of the ice plate from the bottom, is adjustable, giving further temperature control possibilities.

TRIALS Water flow By introducing a dye (malachite green) into the pre-cooling tank the water flow pattern in the experimental tank could be observed. This was done at flow rates of 1 liter min -1 and 2 liters min -1. In both cases the water in the experimental tank became homogeneously colored within a few minutes. The color also disappeared evenly, leaving no 'dead' areas of colored water. It was therefore concluded that the flow pattern through the tank was satisfactory. This was confirmed by taking temperatures at different positions in the tank, when no temperature differences were found. Temperature control Figure 2 shows the daily water temperatures in three experimental tanks during a period of six weeks. The tanks were assigned different temperature regimes which involved gradual cooling, constant low temperature close to OC, and sudden warming followed by gradual cooling. The programs for these regimes are shown by the solid line on the graph. As can be seen from the figure the tank in which the ice plate was used held a temperature of 0-1C throughout the entire six

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G. Sivertsen, J. C. Wallace, T. G. Heggberget

2.5

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Fig. 2. Daily water temperatures in three experimental tanks during a period of six weeks. Dotted lines indicate observed temperatures, solid lines indicate assigned temperatures.

week period, with no deviation whatsoever being noted. The two remaining tanks also performed fairly satisfactorily, the temperatures never varying by more than 0-2C from those intended.

APPLICATIONS This apparatus was designed for incubation experiments with eggs of Atlantic salmon Salmo salar under various simulated natural and extraordinary Arctic river conditions associated with hydroelectrical development. The results of the first of these experiments are reported elsewhere (Heggberget and Wallace, 1983). It is clear that controlled experiments on the developmental biology of overwintering eggs of any Arctic or cold water species can be performed with the same equipment. In addition, experiments which would normally be carried out over long periods and which for that reason are difficult to conduct in small recirculation systems at very low temperatures can be carried out using this apparatus. Such experiments could include, for example, growth and digestion studies close to the lower temperature limits (?) for these processes. Since only non-corrosive materials have been used in construction, the equipment can also be used with seawater, and the temperature range can therefore be extended below 0C. (Some marine organisms live and feed on the undersurface of sea ice.) The applications of the apparatus encompass, therefore, both basic physiology and Arctic ecology.

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Aquacultural uses include, for example, selection experiments for low temperature growth, development and testing of feed intended for digestion at very low temperatures and the control o f egg development in order to obtain alevins at any desired time. At Tromsq~, hatching of salmon eggs fertilized in early O c t o b e r has been delayed until mid June, and the length o f the yolk sac period can probably be extended until August/September, opening the possibility o f an almost year round supply of fingerlings. This method is possibly a cheaper alternative to those employing light and/or hormones to control spawning in adult fish.

REFERENCE Heggberget, T. G. & Wallace, J. C. (1983). Low temperature incubation of salmon eggs. A pilot experiment. Can. J. Fish. Aquat. Sci., in press.

APPENDIX: T E C H N I C A L DATA Cooling system Hitachi compressor, model R G U 302, capacity 6600 kcal h -1. Coolant glycol and water mixture, freezing point --22C. Coolant r e s e r v o i r 150liters. Circulation p u m p s - Flygt type A H V 25/2, adjustable capacity. Coolant pipes - 1 in pipes with 1 in adjustable valves. Bauser timer (for ice plate control).
N o t e . The capacity of the above equipment is sufficient for six experimental tanks with a through flow of 2 liters min -1 in each and a temperature of 8-10C in the incoming water. The coolant temperature would be set a t - 8C.

Header tank
Plastic tank - inside measurements: length = 1.2 m, breadth = 0.6 m, height = 0.8 m. Volume 570 liters. Intake and outlet pipes are 20 mm PVC. Overflow pipe is 32 mm PVC.

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G. Sivertsen, J, C. Wallace, T. G. Heggberget

Pre-coofing tank
Plastic tank - inside measurements: length = 1.0 m, breadth = 0-5 m, height = 0.8 m. Volume 140 liters. Central drain. Cooling coil 25 mm PEH pipe 20 m long.

Temperature control and stirrer unit

Hetotherm 2 stage, effect 1200/600 W. Contact thermometer range 0-30C. Experimental tank EWOS fish tank type number 2 0 1 9 - inside measurements: length = 1 m, breadth = 1 m, height = 0.45 m. Insulation - 50 mm expanded polystyrene on sides and bottom. Cooling c o i l - 25 mm PEH pipe, 20 m long.

Liquid contact parts

The perforated b o t t o m plate in the experimental tank is o f seawaterresistant aluminium. The stirrer and heating coil o f the Hetotherm unit are of stainless steel, and the temperature sensors are embedded in epoxy resin. All other parts of the equipment which contact the water are o f plastic.