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North American Journal of Fisheries Management 27:932935, 2007 Copyright by the American Fisheries Society 2007 DOI: 10.1577/M06-066.1

[Management Brief]

A Method for Determining Stomach Fullness for Planktivorous Fishes

QUINTON E. PHELPS*1
Department of Wildlife and Fisheries Sciences, South Dakota State University, Box 2140B, Northern Plains Biostress Laboratory 138, Brookings, South Dakota 57007, USA

KIPP A. POWELL

AND

STEVEN R. CHIPPS

U.S. Geological Survey, South Dakota Cooperative Fish and Wildlife Research Unit,2 Department of Wildlife and Fisheries Sciences, South Dakota State University, Box 2140B, Northern Plains Biostress Laboratory 138, Brookings, South Dakota 57007, USA

DAVID W. WILLIS
Department of Wildlife and Fisheries Sciences, South Dakota State University, Box 2140B, Northern Plains Biostress Laboratory 138, Brookings, South Dakota 57007, USA Abstract.Mean stomach fullness provides a useful index for quantifying fish diets. However, estimating stomach fullness for planktivorous fishes can be time-consuming and prone to error because of small prey and unidentifiable remains. In this study we developed a predictive equation for estimating the stomach volume of yellow perch Perca flavescens as a function of total length (TL). We then used an optical plankton counter (OPC) to estimate the biovolume of invertebrate prey consumed. The OPC quickly estimated the digital size and abundance of zooplankton prey, which can then be converted to estimates of prey biovolume. Stomach volume (V [mm3]) for yellow perch (113 279 mm TL) was significantly related to body size (L [mm]) and was estimated as V 3 3 107  L2.96. Using the OPC, yellow perch stomach contents (99% Daphnia pulex) were converted to prey biovolume (mm3) and then divided by stomach volume (mm3) to estimate stomach fullness (%). This approach provided reasonable estimates of stomach fullness ranging from 3% to 85% (mean 21%). Although the initial cost for the OPC equipment is relatively high, this method provided substantial time and labor savings compared with traditional approaches for quantifying zooplankton abundance and biomass (e.g., microscopic identification and enumeration and lengthmass conversions). Similarly, the OPC can be used to estimate the abundance and biomass of freshwater zooplankton, thus reducing the time and costs associated with traditional plankton analyses. The approach is limited, however, in cases where very small prey (,250 lm) are a dominant proportion of the sample because of the potential errors involved in detecting and estimating the biovolumes of small particles. *Corresponding author: qphelps@siu.edu 1 Present address: Fisheries and Illinois Aquaculture Center and Department of Zoology, Southern Illinois University, Life Science II, Room 173, Mailcode 6511, Carbondale, Illinois 62901, USA. 2 The South Dakota Cooperative Fish and Wildlife Research Unit is jointly supported by the U.S. Geological Survey, South Dakota State University, South Dakota Department of Game, Fish & Parks, and the Wildlife Management Institute. Received February 13, 2006; accepted January 10, 2007 Published online August 2, 2007

Accurate quantification of fish diets is an important aspect of fisheries management. Traditionally, prey items removed from fish stomachs are identified, counted, and measured for mass or volume. While this approach works well for larger prey such as fish or macroinvertebrates, estimating the abundance, biomass, or volume of zooplankton prey can be time-consuming. Moreover, estimates of zooplankton biomass in fish diets can be compounded by error associated with subsampling and conversions used to estimate zooplankton mass. An optical plankton counter (OPC) developed for automated counting and sizing of zooplankton has been widely used in quantifying plankton biomass in marine environments (Wieland et al. 1997). Recent application of OPC technology to freshwater zooplankton indicated significant correlations between OPC and field-derived estimates of freshwater zooplankton biomass (Sprules et al. 1998). Compared with traditional methods for calculating mass and biovolume for freshwater zooplankton (Wetzel and Likens 1991), the OPC has the potential to provide a new, rapid means for assessing zooplankton biomass from fish diets. A variety of approaches have been used to estimate the stomach volume of fishes, including subjective visual estimates (Hynes 1950), maximum observed prey volume among different fish size categories (Knight and Margraf 1982), and injection of water or air into empty stomachs (Burley and Vigg 1989). Estimates of stomach volume based on maximum observed prey volume provide logical measures, but require large sample sizes across a range of fish lengths. Satiation of the stomach can require fewer fish, but must be performed under artificial settings that may affect the appetite and feeding level of fish. The objective of this study was to assess the usefulness of OPC technology for estimating zoo-

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FIGURE 1.Diagram of the system using an optical plankton counter (OPC) to estimate the biovolume of zooplankton prey from the stomachs of yellow perch collected at Waubay Lake, South Dakota.

plankton biovolume and to use these measures to calculate stomach fullness for yellow perch Perca flavescens. Stomach fullness is a useful diet measure that is correlated with other diet indices such as total prey mass and prey caloric contribution (Pope et al. 2001). To calculate stomach fullness, information is needed on stomach volume of the predator and total biovolume of prey in the stomach. Stomach fullness is then calculated as the ratio of total prey volume to predator stomach volume, thereby providing a diet index that accounts for fish size (Knight and Margraf 1982). Methods Estimating stomach volume.To measure the stomach volumes of yellow perch, we used a modification of the injection technique described by Burley and Vigg (1989). Yellow perch (113279 mm total length [TL]; n 18) were collected through a hole in the ice from East 81 Lake (Kingsbury County), South Dakota, transported to the laboratory, identified to sex, and measured for TL (mm) before removal of stomach contents. Stomachs were excised from each fish and then contents were flushed with water from a 50-mL syringe. After flushing, stomachs were clamped at the pyloric valve (Bond 1979) and liquid epoxy was slowly injected through the anterior end (i.e., posterior end of esophagus) until the entire stomach was distended; the esophagus was then clamped as close to the stomach as possible. After 24 h of drying, stomach linings were carefully scraped away and the epoxy plugs were measured by water displacement to the nearest 0.5 mL. Stomach volumes were then related to fish TL using log10log10 linear regression analysis. Estimating zooplankton biovolume.We collected 40 yellow perch (203267 mm TL) by angling from

Waubay Lake in northeastern South Dakota in February 2002. Yellow perch were collected through a hole in the ice, transported to the laboratory, and examined to determine gender and TL before removal of stomach contents. However, only 55% of these yellow perch (n 22) contained food items; hence, estimates of zooplankton biovolume were based on these fish. Stomachs were excised at the junction of the esophagus and pyloric valve as previously described, and contents were removed by flushing the stomach with distilled water. Stomach contents were then frozen in distilled water until analysis (,2 weeks) to reduce distortion and degradation of zooplankton prey. The freezing process appeared to have little influence on zooplankton degradation; most zooplankton were fully intact after thawing from the ice before processing. The stomach contents of individual fish were processed using an OPC (Model 1 L) connected to a laptop computer (Figure 1). The system was configured as a recirculating system following the manufacturers guidelines (MacKay 1996; Focal Technologies Corporation 1999). To facilitate flow through the OPC unit, 35 drops of a wetting agent (i.e., detergent) were added to distilled water heated to 308C. Water in the recirculating system was pumped from a 20-L holding tank to a 1.3-L reservoir fitted over the OPC. Zooplankton samples from fish diets were added to a beaker and brought to 250 mL with distilled water. Diluted samples were then slowly (;2 min/sample) added to the 1.3-L reservoir while the system was circulating water and allowed to flow by gravity through the OPC before reaching the main reservoir. Samples were then collected in a bucket lined with a 63-lm mesh screen. Before processing any sample, the OPC system was allowed to equilibrate to eliminate air bubbles. The collecting bucket was emptied after each sample and water in the recirculating system was changed after five to eight samples to maintain a water temperature of 30 6 28C. Following OPC processing all stomach contents were combined into a 500-mL beaker and 10% of the sample was examined under a dissecting microscope to quantify zooplankton composition. To estimate the size distribution, the digital size of the zooplankton passing through the OPC was converted to an equivalent circular diameter (ECD) that approximated the diameter of a circular disk blocking the same amount of light as the organism (Sprules et al. 1998; Focal Technologies Corporation 1999). The ECD values generated from the OPC were then converted to volumes (V) using the equation V P ECD3 3 6 1:7689

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TABLE 1.Total length, food volume, maximum stomach volume, and percent stomach fullness for yellow perch (n 22) from Waubay Lake, South Dakota. Means (SEs) are shown in the last row.
Length (mm) 203 204 209 215 215 216 216 217 221 228 231 234 234 240 241 245 247 255 258 265 265 267 234 (4.3) Food volume (mm3) 0.28 1.02 0.07 0.07 0.24 0.61 0.37 0.33 0.63 0.73 1.76 0.54 0.31 0.36 0.84 0.82 0.32 0.16 0.22 0.68 0.21 3.70 0.65 (0.17) Stomach volume (mm3) 1.94 1.97 2.12 2.30 2.30 2.33 2.33 2.37 2.50 2.74 2.85 2.96 2.96 3.19 3.23 3.39 3.47 3.81 3.94 4.27 4.27 4.36 2.98 (0.16) Stomach fullness (%) 14.4 51.8 3.3 3.0 10.4 26.2 15.9 13.9 25.2 26.6 61.8 18.2 10.5 11.3 26.0 24.2 9.2 4.2 5.6 15.9 4.9 84.9 21.2 (4.3)

FIGURE 2.Maximum stomach volume as a function of total length for yellow perch (n 18) from East 81 Lake, South Dakota. See text for details.

(Sprules et al. 1998). Total biovolume generated by the OPC for each sample was divided by the estimated stomach volume of the fish to provide a measure of percentage stomach fullness. Results and Discussion The maximum stomach volume for yellow perch was predicted from the equation V 3 3 107  L2:96 ; where V is the empirical stomach volume (mm3) and L is the empirical fish TL (mm) (log10log10 leastsquares regression; n 18; r2 0.96, P , 0.001; Figure 2). Because bursting capacity was not measured, our approach probably underestimated true maximum stomach distension. Nonetheless, our equation predicted stomach volumes (as a proportion of body size) similar to those reported for other fish species (Knight and Margraf 1982; Pope et al. 2001). A composite sample of stomach contents revealed that Daphnia pulex was the predominant prey item (99%) in yellow perch collected during winter 2002. Ceriodaphnia spp. and adult copepods were also present but represented less than 1% of total zooplankton composition. Zooplankton biovolumes, estimated from the OPC, ranged from 0.07 to 3.7 mm3. Dividing zooplankton biovolume by stomach volume provided reasonable estimates of stomach fullness that ranged from 3 to 85% (mean 21%; Table 1). The OPC provided a rapid analysis of zooplankton biovolumes; the average processing time was about 10 min/fish. Because the OPC does not distinguish between zooplankton species, however, prey composition must be quantified by means of traditional

approaches. Moreover, because the minimum detection limit of the OPC is about 250 lm, it does not detect most rotifers or small copepod nauplii (Sprules et al. 1998). If these taxa are abundant in the diet, then the OPC would probably underestimate true prey biomass. Similarly, small pieces (,250 lm) of zooplankton remains may be underestimated (i.e., not counted) by the OPC; although not evaluated here, this may influence total prey volume of zooplankton removed from fish stomachs. Often times, diet items removed from fish stomachs are in various stages of degradation or digestion. To reduce these problems, we suggest using only prey items found in the stomachs of fishes and not in the intestines. We also recommend that subsamples of fish diets be taken and examined under the microscope, which will allow evaluation of species composition as well as relative abundance of zooplankton remains. Although the shapes of freshwater zooplankton are variable, Sprules et al. (1998) showed that the use of a single geometric model (oblate spheroid) provided accurate estimates of biovolume for mixed zooplankton assemblages. Using mixed assemblages of freshwater zooplankton, they demonstrated that size distribution of zooplankton measured using the OPC were remarkably similar to actual measurements determined from the same samples with a microscope (Sprules et al. 1998). Although comparisons between traditional

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approaches (e.g., microscopy) and OPC measurements help validate OPC-based estimates, traditional approaches can be influenced by sampling and measurement error, thus affecting the accuracy of biovolume estimates. For this reason true validation of OPC-based measurements would be difficult to obtain. Nonetheless, when combined with information on stomach volume, the OPC approach provided quick and reasonable estimates of stomach fullness for yellow perch feeding on cladoceran prey. Depending on the questions being addressed, stomach fullness can provide a useful measure for diet studies; Pope et al. (2001) demonstrated that mean stomach fullness was highly correlated with caloric-based indices that require more time and information to calculate. Moreover, the OPC can be used to estimate abundance and biomass of freshwater zooplankton from field samples, thereby providing multiple uses that reduce the time and costs associated with traditional plankton analyses. References
Bond, C. E. 1979. Biology of fishes, 2nd edition. Saunders, Orlando, Florida. Burley, C. C., and S. Vigg. 1989. A method for direct

measurement of the maximum volume of fish stomachs or digestive tracts. Journal of Fish Biology 34:707714. Focal Technologies Corporation. 1999. Optical plankton counter: users guide. Focal Technologies Corporation, Blacksburg, Virginia. Hynes, H. B. N. 1950. The food of freshwater sticklebacks (Gasterosteus aculeatus and Pygosteus pungitius) with a review of methods used in studies of the food of fishes. Journal of Animal Ecology 19:3558. Knight, R. L., and F. J. Margraf. 1982. Estimating stomach fullness in fishes. North American Journal of Fisheries Management 2:413414. MacKay, I. 1996. Using the OPC 1L laboratory unit: application note no. 4. Focal Technologies Corporation, Dartmouth, Nova Scotia. Pope, K. L., M. L. Brown, W. G. Duffy, and P. H. Michaletz. 2001. A caloric-based evaluation of diet indices for largemouth bass. Environmental Biology of Fishes 61:329339. Sprules, W. G., A. W. Herman, and J. D. Stockwell. 1998. Calibration of an optical plankton counter for use in freshwater. Limnology and Oceanography 43:726733. Wetzel, R. G., and G. E. Likens. 1991. Limnological analyses. Springer-Verlag, NewYork. Wieland, K., D. Peterson, and D. Schnack. 1997. Estimates of zooplankton abundance and size distribution with the optical plankton counter (OPC). Archive of Fishery Marine Research 45:271280.