Aquacultural Engineering 4 (1985) 85-92

Temperature and Size Effects on the Accuracy of Estimating Postlarval Shrimp Populations M.P. Hardin
Cedar Bayou Research Laboratory, PO Box 1396, Baytown, Texas 77520, USA

D.L. Hutchins, G.W. Chamberlain

Texas Agricultural Extension Service and Experiment Station, Texas A&M University, PO Drawer Q, Port Aransas, Texas 78373, USA

and D.V. Aldrich

Department of Wildlife and Fisheries Sciences, Texas A&M University, Building 11, Fort Crockett, Galveston, Texas 77550, USA
A BS TRA CT The common practice in estimating postlarval penaeM shrimp populations for stocking ponds is volumetric subsampling with subsequent extrapolation. This proeedure can lead to considerable undercounting, which, if not recognized, will result in overstocking with accompanying underfeeding and depressed growth. Postlarval blue shrimp Penaeus stylirostris counts derived from volumetric subsampling in a container of known volume and shrimp density were checked for accuracy. Counts were made o f three sizes o f shrimp (4, 6 and 10.5 mm total lengths) at four temperatures (18, 22, 26 and 30C). Water temperature and shrimp size both had highly significant (p < 0.0001) effects on the mean catch, which tended to decrease with increasing shrimp sizes and water temperatures. The population estimates ranged from 100% of the actual population for 4 mm postlarvae at 18C to 57%for 10.5 mm postlarvae at 30C.

INTRODUCTION An accurate technique of estimating postlarval shrimp populations prior to stocking ponds is of practical interest to aquaculturists in both research and commercial production. For example, stocking density and percent survival in pond experiments cannot be calculated without an accurate idea of the initial 85 Aquacultural Engineering 0144-8609/85/$03.30 Elsevier Applied Science Publishers Ltd, England, 1985. Printed in Great Britain

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M. P. Hardin, D. L. Hutchins, G. I. Chamberlain, D. V. Aldrich

population. Further, the large quantities of costly postlarvae used in commercial culture create a need for a quick and accurate method of estimating their numbers. In the past, postlarvae have either been counted entirely by hand or the population has been calculated volumetrically as a product of the estimated shrimp density and the water volume of their container. For the latter technique, shrimp density is derived from the mean number caught in several small-volume aliquots taken from a highly concentrated population. These small volume aliquots have commonly been collected with either a beaker or, more recently, a spring operated pipette. Some pond studies that have employed volumetric estimation of initial populations have given final shrimp survival figures significantly greater than 100% (Berry, 1976; Trimble, 1980; Chamberlain et al., 1981). Nonetheless, in both research facilities and commercial hatcheries there is routine use of the volumetric technique for counting large numbers of postlarvae (Mock and Murphy, 1970; Parker and Holcomb, 1973; Philip Boeing, SEMACUA, Guayaquil Ecuador, personal communication; Edward D. Scura, Aquatic Farms, Kaneohe Hawaii, personal communication). Also, most major contractors for purchase of wild-caught penaeid postlarvae used in the extensive mariculture industry in Ecuador accept volumetrically derived population estimates (although such estimates are generally believed to be chronically low). Hand counting the entire population has the advantage of complete accuracy, but is a tremendous investment of time and labor and may incur greater handling mortality. For these reasons its application has been limited to situations involving low stocking numbers (Willis and Berrigan, 1977; Furness, 1978). The purpose of this study was to test the accuracy of volumetric counting under conservative circumstances and to determine conditions for maximizing accuracy.

MATERIALS AND METHODS Approximately 10000 first day postlarvae of blue shrimp Penaeus stylirostris were obtained from the Texas A&M Mariculture Research Laboratory. The shrimp were transported to the Cedar Bayou Mariculture Research Laboratory, where the experiments were performed.

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The initial total mean length of the shrimp was 4 mm. All measurements were made to the nearest 0.5 mm under 10X magnification from the tip o f the rostrum to the tip of the telson. For each experiment 500 postlarvae were counted individually after being caught on handsized pieces of 1.5 mm mesh screen. These were released into a l-liter plastic beaker containing 500 ml o f seawater, resulting in a density of 1 postlarva m1-1. The water temperature was adjusted by partially immersing the beaker in warm or cold water baths. The beaker was then inserted into a styrofoam sleeve to help maintain the experimental temperature. Water temperature and volume were checked approximately every 3-5 min throughout each experiment and corrected as needed. Pure oxygen was bubbled continuously in the beaker. A Hensen-Stempel automatic pipette (Wildlife Supply Co., Saginaw, Michigan) was used to subsample. The pipette takes a 10 ml volume of water by means o f a spring-operated plunger. Operation o f the pipette is very quick, requiring less than 1 s to take an aliquot. This pipette has been routinely used by Texas A&M University staff to subsample penaeid eggs and nauplii, but its efficiency with postlarvae was unknown. Each aliquot was first discharged into a plastic dish to count and record the number of captured postlarvae. Healthy shrimp were then returned to the beaker. Shrimp mutilated by the pipette (approximately 20% of those captured) were replaced from reserves. To achieve an even distribution of postlarvae in the beaker, the water was vigorously stirred immediately prior to taking each 10 ml subsample. To prevent concentration of the postlarvae in the middle o f the beaker, stirring was done in a side-to-side then front-to-back manner. A tightly rolled tied plastic bag served as a soft, stiff stirring rod. 30 aliquots were taken for each of the temperatures (18, 22, 26 and 30C). Fresh batches of 500 postlarvae were used at each temperature. All o f the postlarvae were then divided between two 380-liter aquaria and fed brine shrimp nauplii daily. After 1 week, mean length had increased to 6 ram. Following methods described above, the experiment was repeated at the same four temperatures. Yet another repetition at these temperatures was performed after a second week of growth in the aquaria had increased mean shrimp length to 10.5 mm. Means and standard deviations were calculated for the number caught per aliquot at each temperature and size. Two-way analysis of

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M. P. Hardin, D. L. Hutchins, G. W. Chamberlain, D. V. Aldrich

variance (ANOVA) was performed on the data to check for significant differences among treatment levels. This was followed by Duncan's multiple range test (DMR) to identify specific significant differences among means. Catch variabilities among treatments with different means were compared using coefficients of variation (Sokal and Rohlf, 1969).

RESULTS A catch o f 1 postlarva m1-1, or 10 per aliquot was averaged at the 18C temperature with the 4 mm length (Fig. 1). However, mean catch decreased with increasing size and temperature. Catch was least with 10.5 mm lengths at 30C. ANOVA indicated highly significant differences in means between levels of both factors (for temperature: F = 10.48, degrees o f freedom = 3, p < 0.0001 ; for length: F = 40.24, degrees of freedom = 2, p < 0 . 0 0 0 1 ) . Specific significant differences among means, as isolated by DMR, generally occurred between temperatures o f 18 and 22C and sizes of 4-6 mm (Fig. 1).

IMEAN CATCH

Fig. 1.

Mean catch of 4.0-10.5 mm length postlarvae at 18, 22, 26 and 30C. Means having the same letter are not significantly different (p < 0.05).

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An accurate estimate of postlarval density was thus obtained with 4 mm size postlarvae at all four temperatures. However, density was underestimated with larger postlarvae, particularly at the higher temperatures. Specifically, at 18C, the mean catch derived from repeated subsampling o f 6 and 10.5 mm postlarvae underestimated the actual density by 16%. At 22-30C the indicated density for the same size group was 31-43% less (respectively) than the actual density. The latter finding is important, since the temperature and size ranges of 22-30C and 6-10.5 mm are those most likely to be encountered by culturists when stocking ponds. A 95% confidence interval about the mean catch (N = 180) for these ranges was established as 6-47 0.37 (64.7 + 3.7% accurate). Coefficients of variation were large, ranging from 22.0 to 47.3%, with lower values more common among the smallest size of postlarvae (Fig. 2).

DISCUSSION Reduced catches are probably due to the shrimps' avoidance of the pipette. The postlarvae remained active throughout the experiment and

tCOEFFICIENT OF VARIATION

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Fig. 2.

Coefficients of variation for the mean catches (Fig. 1) of three sizes of postlarvae at four temperatures.

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M. P. Hardin, D. L. Hutchins, G. W. Chamberlain, D. V. AMrich

were observed to jump back from any object thrust into their immediate vicinity. Reaction times would be expected to decrease with increased temperature, since shrimp are poikilotherms. Increased size would further enhance their ability to avoid something as limited in effective range as the pipette. The high variation from aliquot to aliquot in the number of postlarvae captured is typical for the volumetric counting technique. This indicates that patchy distributions persist in a contained population of postlarvae, even after vigorous mixing. The practical significance is that if thoroughly mixed populations of postlarvae were not obtained under our conservative, small-scale conditions, then good mixing would be even more difficult when sampling from large containers. In summary, at temperatures ranging from 18 to 30C, volumetric estimation is a fairly accurate technique for counting live postlarval shrimp that do not exceed 4 mm in length. Density estimates for the larger postlarvae (such as those used to stock ponds) are underestimated by a relatively standard factor dependent on temperature and size. Data presented in Fig. 1 represent an index of the extent of avoidance capability of postlarval shrimp with increasing size and temperature. This phenomenon may often escape detection, since postlarval mortalities in ponds are commonly very high. If final populations are reduced to levels less than stocked populations, no initial miscounting would be suspected. A solution to underestimation could be to: lower water temperature prior to sampling; use a larger volume subsampling device; or revise the population estimate upward using experimental correction factors. An example of the latter option based on 64.7% accuracy of the present experiment implies that the real population is 100/64.7 = 1.55 times the estimated population. Chilling the water prior to subsampling would be cumbersome in execution, stressful to the postlarvae, and yield an estimate still lacking in complete accuracy. The use of a larger volume sampling device, such as a 25 or 50 ml pipette, is currently under investigation by Texas A&M personnel. However, this modification would increase the number of postlarvae per aliquot, in turn increasing the time required to count a big population. Further, even the use of a 400-ml beaker in the Trimble (1980) study did not prevent a substantial underestimation of stocking density. Thus, some manner of correction factor seems necessary for the volumetric counting technique.

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The correction factor of 1.55 from this study was used to estimate the stocking populations o f P. stylirostris postlarvae used in the 1980 Cedar Bayou pond experiments (Hardin, 1981). Harvest populations in these experiments ranged from 71 to 100% of the corrected stock population (implying that uncorrected estimates would have yielded survivals greater than 100%). The use o f different species, sizes, techniques, equipment, etc., may require experimental calculation of different correction factors.

ACKNOWLEDGEMENTS This research was funded by the Houston Lighting & Power Company through the Department of Wildlife and Fisheries Sciences and Texas Agricultural Experiment Station Project 1869-2339.

REFERENCES Berry, R. (1976). Pond study crustaceans. In: The Effects on Selected Organisms of Water Passing Through the Cedar Bayou Generating Station, Texas Agricultural Experiment Station Project Report for 1975, pp. VII-1-VII-I 3. Chamberlain, G. W,, Hutchins, D. L. & Lawrence, A. L. (1981). Mono- and polyculture of Penaeus vannamei and P. stylirostris at differing percentages in experimental ponds. Proceedings WorldMariculture Society, 12 (1), 251-70. Furness, G. N. (1978). Growth, survival, distribution and population estimates of penaeid shrimp in fertilized and unfertilized brackish water ponds receiving thermal effluents. MS Thesis, Department of Wildlife and Fisheries Sciences, Texas A&M University. Hardin, M. P. (1981). Considerations of diet, stocking density, distribution, population estimation and economics in the pond culture of blue shrimp, Penaeus stylirostris (Stimpson). MS Thesis, Department of Wildlife and Fisheries Sciences, Texas A&M University, 155 pages. Mock, C. R. & Murphy, A. (1970). Techniques of raising penaeid shrimp from egg to postlarvae. Proceedings Worm Mariculture SoeieO,, 1, 143-56. Parker, J. C. & Holcomb, H. W. Jr. (1973). Growth and production of brown and white shrimp (Penaeus aztecus and P. setiferus) from experimental ponds in Brazoria and Orange counties Texas. Proceedings Worm Mariculture Socie~. , 4, 215-34. Sokal, R. R. & Rohlf, F. J. (1969). Biometry, W. H. Freeman and Co., San Francisco.

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Trimble, W. C. (1980). Production trials for monoculture and polyculture of white shrimp (Penaeus vannamei) or blue shrimp (Penaeus stylirostris) with Florida pompano (Trachinotus carolinus) in Alabama, 1978-1979. Proceedings World Mariculture Society, 11,44-59. Willis, S. & Berrigan, M. (1977). Effects of stocking size and density on growth and survival of Macrobrachium rosenbergii in ponds Proceedings World Mariculture Society, 8, 251-64.