Aquacultural Engineering 1 (1982) 63-70

DESIGN AND DEVELOPMENT OF A FLOWING BED REACTOR FOR BRINE SHRIMP CULTURE

D. E. BRUNE

Department of Agricultural Engineering, University of California, Davis 95616, USA

ABSTRACT

An automatic self-cleansing high density brine shrimp reactor has been designed and tested, and the concept appears successful Initial experiments suggest that extraordinarily high densities o f brine shrimp may be maintained in such a system. The concept o f a flowing film o f glass beads appears to be a very useful technique in scouring waste material from animal culture tanks. The concept is particularly useful for culture o f small marine herbivores in continuous culture. The bead flow rapidly removes waste material while allowing for water passage, y e t completely containing the animals. Further design improvement and development should result in a high density brine shrimp culture system which is both simple and inexpensive to build, as well as reliable and cost effective to operate, and capable o f efficient algal cell removal from culture water.

INTRODUCTION

The tremendous photosynthetic productivity of mass algal culture has for years excited both scientists and entrepreneurs alike. Yet after 30 years of experimentation, construction and operation of laboratory prototypes and pilot plants, fundamental problems remain to be solved before algal culture can be seen as a viable and cost effective means of protein production. Three major problems have prevented the widespread utilization of algal culture as a protein source. First, large scale algal cultures are often seen to become dominated by undesirable 'weed' algae. Secondly, even if a desired algal species can be maintained, many of the fast growing unicellular algae are poorly digestible by monogastric animals. And finally, unicellular algal culture has proved to be difficult to harvest economically. 63
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A variety of recent studies have attempted to come to grips with these problems. Perhaps the most promising approach is that advocated by Ryther et al. (1975) in which various f'dter feeding organisms are used to glean algal cells from the culture medium and convert them into a desirable and readily harvested animal protein source. Ryther et al. (1975) and Goldman and Ryther (1975) have demonstrated the use of secondary effluent from an activated sludge waste water plant as a fertilizer for culture of marine phytoplankton. With the addition of from 20% to 50% sewage to marine ponds, it was found that a continuous high rate production of marine diatoms could be maintained. The diatoms were in turn to be used as a feed for various shellfish. The concept of using marine herbivores apparently solves the problem of algal harvest, with the algal culture being converted into a useful end product. The major disappointment of field culture studies has been the inability to maintain cultures of desirable algal species. The marine cultures are often observed to become dominated by certain 'weed' (in most of the marine work the diatom Phaeodactylum) algae. Unfortunately Phaeodactylum as well as the other weeds are not utilizable by many of the desired marine filter feeding organisms (i.e. clams, oysters). The solution to this problem lies in the selection of a filter feeding animal capable of growth on these commonly occurring 'weeds'. One such animal, the brine shrimp Artemia salina, has recently received much attention. This animal seems ideally suited for algal biological conversion systems. The ability of this animal to utilize many of the fast growing dominating (Takuno, 1967) algae combined with its rapid growth rate (Reeve, 1963), high protein content, and high tolerance of environmental stress, suggests that this animal may be the key to finally harnessing the potential productivity of algal culture. It is the intent of this presentation to describe recent research which was undertaken in an effort to develop further the technology of high density brine shrimp culture.

DESIGN METHODS AND MATERIALS

Previous culture methods A recirculating shrimp culture system was described by Mock et al. (1973). This system consists primarily of a concrete hatchery raceway with a center drain. Waste removal in this system was accomplished via the center outlet which was equipped with either a slotted pipe or plastic filter screen to prevent animal loss. Water circulation was maintained by the use of multiple air lift pumps. Sorgeloos et al. (1979) used a modified version of Mock's raceway system for culturing brine shrimp. Experiments were conducted in which brine shrimp were grown for periods of two weeks or more by operating the raceways as recirculating batch systems. The feed used consisted of additions of various finely divided particulate organics. In early experiments, no attempt was made to remove particulate waste production from the cultures. In more recent works, such attempts were made

FLOWING BED REACTOR FOR BRINE SHRIMP CULTURE

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in hopes of improving and maintaining culture water quality. To date the most successful results have been obtained by the use of cross flow screens to separate animal biomass from particulate waste material. On a larger scale, Milligan et aL (1979) report on the use of a dual tank system which utifizes a central 'high' density brine shrimp tank, located within a lower density larger volume tank. The algal feed source enters into the smaller central tank. At the outline of the smaller tank an air-lift pump returns the majority of the brine shrimp to the small central tank. However, a certain portion of the animals are always escaping to the larger outer tank; thus, this system does not effect a true separation, but rather acts as a concentrator of brine shrimp. As a result, the internal tank is limited to a rather low total animal density of only 300-400 animals/liter. Tobias et al. (1979) described the use of modified plastic aquaria for culturing brine shrimp on algae. In their system the brine shrimp were separated from the outflow by use of fine mesh nitex screens. Fine mesh screens were also used on 190 liter raceway cultures, the size of the screens being manually changed as the brine shrimp grew. Clogging of the screens by particulate waste was a problem resulting in animals being lost from the cultures when they overflowed. The previous methods used to retain small herbivores such as brine shrimp in culture vessels fall into two types: the concentrator system such as the DOW system, and the use of fine mesh screens. Both methods have disadvantages. The concentrating method appears limited to low biomass levels, thereby requiring very large culture volumes, which in turn implies a large capital investment. The use of fine mesh screens allows for a much higher animal biomass, but clogging problems and the need to change screen size as the animals grow makes this an awkward design for potential use in large scale automatic cultures. Furthermore, it may be desirable that such a system be capable of operating with a random distribution of animal sizes. The use of screens does not meet this requirement. A system is needed that can: 1. Completely retain the brine shrimp at all stages of growth at high density. 2. Allow for rapid and complete particulate waste removal. 3. Allow for a high rate of water passage to deliver the needed algal suspension. Two design concepts are of particular value in meeting these requirements. First the concept of the simple aquarium subgravel filter, a concept utilized by Farrar (1977) for culture of oyster larvae; although effective in removing wastes, the system as presently utilized cannot handle heavy waste loads. However, the second concept described by Sandifier et al. (1974), the use of a tank with an inclined floor to achieve movement of waste material to an outline, provides the answer to the loading problem. These two concepts were used as the basis of a new system; the design was formulated and a small prototype reactor was built and tested.
Trial runs with adult brine shrimp

The first experimental unit (Fig. 1) consisted of a 12 x 12 x 8 in (30 x 30 x 20 cm) plexiglas tank with baffles 1 in (2.5 cm) from the ends, and open ~ in (1.905 cm) at

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Fig. 1.

Original flowing bed reactor.

the bottom. The floor of this vessel was filled with Ferro microbeads in the size range of 500-710 #m. This size grouping was found to be most effective in containing the adult Artemia, yet providing high porosity for water passage. Two air-water lift pumps ran along the b o t t o m of the tank under the bed floor. These pumps consisted of two ~in (1.905 cm) plexiglas tubes with a 90 bend at the rear baffle. The tank was positioned on a wooden support with a provision for changing the angle of incline. To begin operation, the tank was filled with a synthetic sea water mixture ('Instant Ocean') and enough beads were added to achieve the ~in (1.905.cm) bead depth. Next the tank was inclined until the angle of repose of submerged beads was achieved (i.e. the point at which they begin to flow). This angle was found to be 30 for the 500-710/Jm size group. Air flow to the air-water lift was started at this point. The animal compartment (the center volume) was then loaded with 8000 adult Artemia per liter. For the adult Artemia runs, a suspension of whole wheat flour was fed in at a rate which gave a hydraulic retention time of about 8 h. The operation of the system was as follows: in the front compartment, the airwater lift pumps would continually lift a slurry of beads and particulate waste to the

FLOWING BED REACTOR

FOR BRINE SHRIMP CULTURE

67

rear compartment. In this compartment the lighter waste particulates would separate from the beads as the heavy beads settled rapidly to the bottom with the waste material remaining in the water column as a result of the bubbling action of the airwater lifts. As the beads travel across the floor of the tank, particulate wastes settle on and into the bed. At the front baffle, the waste-loaded bed slumps under the plate to be removed once again by the air-water lift. The animals; however, respond to the flowing floor as if it were a solid surface and remain behind. The cleansed beads continually re-enter under the rear baffle. In these initial experiments, the tank was operated over a period of 72 h. With a bed travel time across the bottom of approximately 30 rain, the cleansing action was found to be more than sufficient. The flowing bed rapidly removed wastes from the tank, while confinement of the adult brine shrimp was complete. The only animals carried through the bed (and to the settling tank) were those which were apparently damaged in handling and transfer. These dead and dying animals were effectively removed from the culture by the flowing bed action. Although this bed size range was quite successful in maintaining a high density of adult brine shrimp, it did not contain the nauplii which were being released by the adult population. At the end of this experimental run, a high density of nauplii were found swarming in the settling tank. This result was carefully noted, as it suggests the possibility of an automatic continuous means to separate nauplii from a brine shrimp brood stock.
Trials runs with Artemia nauplii

A second, larger culture vessel (114 liters) was constructed (Fig. 2) incorporating the same basic principles used in the first reactor. The purpose of this second series of experiments was twofold. First, it was necessary to evaluate a series of bead diameters with regard to nauplii retention and porosity to water flow. In addition, a more rugged apparatus was needed so that the performance of the flowing bed could be evaluated on a longer term basis. In order to study the characteristics of the very small diameter glass beads, it was necessary to modify the tank design to include the use of a supported nitex screen. This screen was included purely as a research tool to allow for an evaluation of the bead flow rate at bead diameters (105-149/3m) giving very low porosity. The angle of inclination in these experimental runs was increased to 45 . At this angle, the bed depth was decreased to less than ~ i n (79.4 mm). The operation remained basically the same as with the original culture device, except that the flowing bed was more of a thin scouring film and the bulk of the water now would exit through the flowing film and across the supporting screen, rather than exiting as a bead-water slurry at the outlet sump. The reactor was set into operation with the air-water lift pumps adjusted to pump a total volume of 3 liters/h each. Of this volume, approximately 60% was beads. The total bead volume in the 30 gallon (136.5 liters) tank was set at 1.6 liters, which gave a bed detention time of 45 min with a floor velocity of 0-5 in (1.27 cm)/s. Aeration for

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D.E. BRUNE
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Modified flowing bed reactor.

the culture was supplied primarily from the action of the air-water lift pump. An additional aeration tube was used in the tank itself for the purpose of mixing the culture in a gentle fashion. The tank was loaded with 145 000 Artemia nauplii per liter, and a flow rate of 18 liter/h of algal suspension was fed into the tank. The alga used in this experiment was Phaeodactylum sp. produced from a 114 liter continuous culture. Influent cell densities averaging 8.5 x 10 s cell/liter were maintained. The reactor performance was monitored for a period of 16 days, primarily to evaluate the effectiveness of the retention of the nauplii. During this period measurements of the brine shrimp biomass, algal cell counts and dissolved oxygen levels were made once each day.
DISCUSSION

Culture behavior The productive capacity of the continuous algal culture was quite limited compared to the extraordinary brine shrimp density being maintained. The brine shrimp biomass

FLOWING

BED REACTOR

FOR

BRINE

SHRIMP

CULTURE

69

outgrew the algal culture by the fourth day. However, during this short period the success of flowing bed in retaining the nauplii was complete and readily apparent. It is interesting to examine the data during the first few days of operation (Fig. 3). The growth of the brine shrimp biomass is reflected in the decline in the dissolved oxygen levels. This growth is also reflected in the rapid decline in effluent algal cell density within the initial 72 h of operation. By day four the culture was already severely food limited as seen in the cell counts; this is further shown in the upturn in culture D.O. on day five, as a result of falling respiration rates.
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Fig. 3. Culture algal cell counts and dissolved oxygen levels during first eight days of operation.

Beginning on day five, a suspension of rice bran was fed into the culture over a 48 h period. The resulting drop in the D.O. was dramatic, vividly illustrating the price that must be paid when attempting to use particulate organics in such systems. In spite of short detention times and rapid particulate uptake rates, organic loading can be expected to impose a heavy oxygen demand on such a system. This observation is important to note in view of the various systems proposed for the utilization of particulate organics by brine shrimp (Sorgeloos, 1979). Because of severe food limitation, a high rate of mortality was observed from day six onward. However, the reactor was allowed to operate to further observe the cleansing action. The system was quite efficient in removing particulate matter, although the importance of an unobstructed bed was noted. Any 'dead spots' would immediately accumulate waste material. The reactor was operated continuously for a total of one month. During this time, the bed remained completely clean. Retention of the Artemia was complete during proper operation.

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Design i m p r o v e m e n t

D.E.

BRUNE

Two potential problem areas were identified: first, it was found that if air flow to the air-water lift pump was even momentarily interrupted, the pump would clog with beads. Secondly, it was found that design of the bead inlet led to unstable operation. The success of the separation was dependent on a volume of beads being retained in the rear inlet box. Occasionally this level would fall and an open communication would be established through which brine shrimp were lost to the settling tank. Design modifications are currently being tested to solve these problems. The apparent solution lies in the development of a low rpm feeding device at the bead inlet and outlet. This device will give control of bead velocity and detention time. With proper selection of blade clearance in the feeder, it has been possible to retain the beads while allowing for water passage, thus giving independent control over the hydraulic detention time and the scouring rate. The development of a feeder also solves the clogging problems with the air lift pumps, since bead flow is restricted to a rate well below the delivery capability of the air lift pumps.

ACKNOWLEDGEMENTS The author wishes to express his appreciation to Patrick Burke for his assistance in developing and constructing the flowing bed models.

REFERENCES Farrar, S. W. (1977). Mortality problems in oyster hatcheries. Fish Farming International, March 1977. Goldman, J. C. & Ryther, J. H. (1975). Nutrient transformation in mass cultures of marine algae. J. Env. Eng. Div., June. Milligan, D. J., Quick, J. A., HiU, S. E., Morris, J. A. & Hover, R. J. (1979). Sequential use of bacteria, algae and brine shrimp to treat industrial wastewater. Syrup. on the Brine Shrimp Artemia salina, Corpus Christi, Texas. Mock, C. R., Neal, R. A. & Salser, B. R. (1973). A closed raceway for the culture of shrimp. Proc. 4th Annual Workshop World Mariculture Society, Monterrey, Mexico, pp. 247-59. Reeve, M. R. (1963). The filter feeding ofArtemia. J. Exp. Biol., 40. Ryther, J. H., Goldman, J. C., Gifford, C. E., Huguenin, J. E., Wing, A. S., Clarner, J. P., Williams, L. D. & Lapointe, B. E. (1975). Physical models of integrated waste recycling. Aquaculture, 5, 163-77. Sandifier, P. A., Zielinski, P. B. & Castro, W. E. (1974). A simple airlift operated tank for closedsystem culture of decapod crustacean larvae. Helgolander wiss Meeresunters, 26, 82-7. Sorgeloos, P. (1979). The culture of Artemia salina on rice bran. lOth Annual Meeting o f World Mariculture Society, Honolulu. Takuno, Hideaki (1967). Rearing experiments of brine shrimp on diatom diet. Bull. Tokai Reg. Fish Res. Lab., No. 52. Tobias, W. J., Sorgeloos, P., Bossuyt, E. & Reels, O. A. (1979). The technical feasibility of mass culturing Artemia salina in the St. Croix artificial upwelling mariculture system, lOth Annual World Marieulture Society Meeting, Honolulu.