x°/EPA United States Environmental Protection Agency Environmental Research Laboratory Narragansett Rl 02882 Research and Development EPA-600/S3-82-017 July 1982 Project Summary Development of Techniques and Methodology for the Laboratory Culture of Striped Bass, Morone saxatilis (Walbaum) Bruce A. Rogers, Deborah T. West in, and Saul B. Saila This summary describes the research undertaken to develop laboratory culture techniques for striped bass (Morone saxatilis) that could be used to provide an adequate supply of various life stages of this important fish species for water quality and hazard evaluation testing. For each of the four life stages defined here (egg, larval, juvenile, and adult) the upper and lower lethal levels where ap- plicable and an approximation of optimum conditions were defined with regard to physical characteristics of the environ- ment including temperature, salinity, dissolved oxygen, light, and turbidity. Satisfactory laboratory diets were de- fined and verified for each life stage. A comprehensive set of procedures was developed and described in a step-by- step manner for use by research person- nel wishing to maintain laboratory pop- ulations of striped bass for physiological and toxicological use. This Project Summary was developed by EPA's Environmental Research Labo- ratory, Narragansett, Rl, to announce key findings of the research project that is fully documented in a separate report of the same title (see Project Report ordering information at back}. Introduction Striped bass, Morone saxatilis, is an important commercial and sport fish species with a center of distribution be- tween the Hudson River and the mouth of Chesapeake Bay. Individuals of this species ascend major rivers to spawn, use coastal estuaries as nursery grounds, and as adults make seasonal migrations along the coast rarely straying more than five miles from the shoreline. Because it passes its entire life cycle in the waters immediately adjacent to the Boston- Washington, D.C., megalopolis, it is subjected to the most intense effects of man-made pollution and environmental alteration. In spite of these abuses, the Atlantic population of striped bass has until recently enjoyed great abundance. Although in the past a considerable amount of research has been done on the culture of the species for stocking into southern reservoirs, no reliable cul- ture methodology has been developed for maintaining all of the life stages of the striped bass in the laboratory where the effects of various pollutants may be determined in physiological studies and bioassay experiments. This study was undertaken to develop a reliable culture protocol for all life stages of the striped bass. Armed with such a protocol, researchers will be in a better position to examine the effects of water borne pollutants on this resilient but vulnerable species. ------- Discussion The striped bass is a desirable candi- date for toxicological investigations in the United States for the following reasons. The species inhabits a wide range and is distributed along all three coasts. It is a commercially and recrea- tionally important species throughout its range. It is also an ecologically important member of the community it inhabits, not only along the coasts, but also within the coastal plain rivers. Its life stages are euryhaline and eurythermal, making them extremely useful in studies to determine sublethal differences in the physiology of toxicants over broad salinity and/or temperature ranges. In addition to being easy to culture, a great deal of the back- ground research on this species has been reported. To date, striped bass culture has been undertaken primarily by federal or state fish hatcheries to stock lakes, reservoirs and impoundments for sport fishing and shad (Dorosoma sp.) control. This work is done almost exclusively in fresh water, either in a hatchery or in ponds (Bonn et al., 1976). The culture methodology recommended in this report for the life history stages of the striped bass, how- ever, utilized sea water wherever possible. This was the case not only because of its availability and cost-effectiveness of use, but primarily to keep disease prob- lems to a minimum. Although some fresh water (to reduce salinity) is needed during spawning and early larval stages, juveniles and adults feed and grow in sea water. Using the methods recommended in this research and summarized below, striped bass life history stages can be cultured which are representative of the species. Figure 1 and Table 1 describe developmental stages of striped bass larvae to metamorphosis. Culture Methods Outlined The outline of recommended methods to secure batches of larvae or juveniles for toxicological studies that follows is based on the details presented in Sec- tions 8-1 1 of the full report available from NTIS. In general, if 20,000 prolarvae are required for studies, then a minimum of 40,000 eggs are needed. This estimate is based on 50% survival, although egg survival varies from 10-20% for artifi- cially spawned to 60-90% for naturally spawned (see Table 2) and fertilized eggs. This survival rate can be increased to 40-50% by using antibiotics in the rearing water. If 2,000 post-larvae are required, then 2,500 prolarvae (80% survival through initial feeding), or 5,000 eggs are required. These estimates are provided as a guide and may vary with broodstock, investigator, facilities and other variables. A. Fertilized eggs can be obtained from natural or artificial spawnings. 1) Artificially spawned eggs re- quire maintenance of broodfish (mature adults) in culture system equipped with temperature, sa- (b) (a) Figure 1. Developmental stages (after Manuseti, 1958) of striped bass larvae to metamorphosis. Refer to Table 1 for further description of stages. ------- Table 1. Developmental Stages Age of Striped Bass, Reared at About 17°C, Unless Otherwise Stated, Through Transformation Length mm TLa Characteristics 25.8 hours after fertilization (4)>> 3.25-4.06 36-48 hours after fertilization (2) 2.5-3.7 51.8 hours after fertilization (4) 3.25-4. 71 1st day after hatching (4) 3.58-5.12 4.23-5.20 2-5th day after hatching (1,2) 4.5-5.2 3rd day after hatching (3) 5.2 (41 4.71-6.23 5.O4-5.77 4th day after hatching (3) 5.8 (4) 5.5-7.5 (live) 5th day after hatching /I) 5.5-5.8 6th day after hatching 13) 6.0 6th- 7th day after hatching (4) 5.5- 7.5 (live) 8th day after hatching (1) 5.8-6.5 (3) 6-9 10-15th day after hatching (2) 7.5 10th day after hatching (3) 9.0 15th day after hatching (1) 10-12.5 18th day after hatching (3) 13.0 20-30th day after hatching (1,2) 10, 12-16 30 days after hatching (4) 13.1-15.4 30-40th day after hatching (2) 15 (stunted) 40 days after hatching (4) 11.9-20.4 40-50th day after hatching (1) 22-35 50- 70th day after hatching (1) 35-45 (2) 20 60-80th day after hatching (2) 25 Hatching completed for eggs at 24 °C. (aft Hatching occurs, (a) Hatching completed for eggs at 18°C. (a) Eyes almost fully pigmented; pigmented ventrally; one-third yolk reabsorbed at 24 °C. Eyes only partially pigmented; yolk slightly reabsorbed at 18°C. Yolk sac partly absorbed, eyes pigmented yellow, black & orange, differentiation of jaws and digestive tract begun, pectoral buds formed fan-like fin, 21 -23 myotomes. (b) Eyes pigmented, jaws developing, pectoral fins become differentiated. Eyes pigmented; mouth pans moving; pigmented ventrally jaw to oil; yolk three-fourths reabsorbed; pectoral buds present at 24 °C. Eyes pigmented; gut differentiated; ventrally pigmented; pectoral buds visible at 18°C. Small chromatophores along ventral edge of entire yolk sac. Yolk absorbed at 24 °C. One-third yolk reabsorbed, commencement of intestinal peristalsis, 23-24 myotomes. Swimming pelagically. (c) Oil globule and yolk nearly absorbed, pigmentation ventrally. (c) Yolk absorbed at 18°C. (d) Teeth on jaws, orange pigment in caudal (heteroceral) area, differentiation of stomach, three-fourths yolk reabsorbed, 25 myotomes. Transition to active pelagic feeding, (d) Second dorsal and anal slightly differentiated, well-developed mouth parts, (d) Yolk sac fully absorbed and no oil globule visible, pectorals only fins visible, teeth visible, generally pigmented on body, (e) Pectorals only fins developed, ready for food. Division of fin fold into three divisions, complete reabsorption of oil globule, single-chamber gas bladder filled with air. Feeding on plankton, (e) Dorsal and anal fin rays well differentiated and rudimentary spines observed, (f) Differentiation of rays in caudal, anal and dorsal fins. First dorsal elements and pelvic fins absent, myotomes correlated with number of vertebrae, (g) Metamorphosis at 24 °C. Soft dorsal, anal and caudal (homocercal) fins well differentiated, spinous and pelvic fins not well developed and well ossified, no stripes visible yet. Initial formation of lateral-line scales (Murawski, 1958). (h) Metamorphosis at 18°C. Differentiation of rays in first dorsal and pectoral fins. Full complement of lateral-line scales by 30 mm (Murawski, 1958). Scales Scales observed for first time, fins except larval pelvic in various stages toward full meristic count, pigmentation stronger. Covered with scales, 3 anal spines and full complement of meristic characters, body covered with melanopores. ------- Table 1. (continued) Age Length mm TL" Characteristics 80-90th day after hatching (1) 50-80 70-100th day after hatching (2) 30 3-4 weeks after hatching 13) 36 Appearance of longitudinal stripes. Meristic counts complete except for fin rays, body pigmentated. Fully developed fins and rays, pigmentation of black dots. ' Total length measured on preserved samples unless otherwise stated. b Numbers in parenthesis refer to source, i.e., (1) Doroshev (1970); (2) Mansueti (1958); (3) Pearson (1938); and (4) Rogers et al. (1977). 0 Letters in parenthesis refer to Figure 11. Table 2. Percent Survival Through Hatching of Striped Bass Eggs from Artificial and Natural Spawnings Incubation Salinity* (O/oo) Artificially induced spawning 0 5 10 15 Incubation Temperature (°C) 16 18 20 58.5 64.3 (561) (280) 1.2 - (249) 19.3 - (165) 31.2 (160) — 7.4 (244) 11.6 (215) 0 21 4.7 (536) 5.4 (185) 11.6 (205) • 0 Natural matured spawning 0 10 15 77.9 (384) 90 (10) 90 (10) 90 (10) 71.0 (473) 71.5 (421) 90 (10) 90 (10) 80 (10) * Percent survival at Q°/oo and 16°C (60°F), 18°C (65°F), and 21 °C (70°F) reported by Shannon (1970). Survivals at the other salinity-temperature combi- nations are results of this study. + ( ) = number of eggs per treatment. Unity, and photoperiod control with subsequent controlled spawning. 2) Naturally spawned eggs may be obtained easily by plankton or neuston net fishing in spawning rivers at the time of spawning (February-May). 3) Collection of naturally spawned eggs insures genetic diversity not available among progeny of a mating under controlled spawning. B. Handling of fertilized eggs to maxi- mize survival and hatching. 1) Eggs collected from plankton tows must be separated before transporting them to rearing containers. 2) Eggs secured from artificial spawnings can be stocked directly into rearing containers at a rate of approximately 100 per liter. 3) Handle eggs only in water, i.e., dip or pipette or siphon. Do not use dip nets. 4) When transferring shipped eggs to rearing containers check that water temperature of two are within 1 °C of each other and that rearing water quality is opti- mum for egg survival (Table 3). 5) Water quality, especially tem- perature, dissolved oxygen and salinity, should be monitored daily and maintained at optimum levels (Table 3). C. Handling of larvae to maximize sur- vival and growth. 1) Easiest method of securing pro- larvae is from eggs on hand that hatch. 2) The recommended larval rearing system is static prepared tank system modified from Houde & Ramsey (1971) and described in greater detail in the full report available from NTIS. 3) Larvae can be stocked at 100 per liter until actively feeding, when densities should be re- duced to approximately 50 per liter. 4) Growing larvae should be graded to nearly equal size fish to reduce cannibalism. 5) The water quality in larval rearing containers should be monitored daily and maintained at optimum level (Table 3). 6) Larvae should be fed 10-20% of their dry body weight at least twice daily beginning about 4-5 days after hatching an approved strain of newly-hatched brine shrimp nauplii. Table 4 lists bio- chemical characteristics of some life diets for larval bass. 7) Growth rates at various temper- atures are detailed in Figure 2. 8) As larvae reach metamorphosis other foods such as ground squid or prepared diets (moist pellets) can be added to adult brine shrimp. ------- Table 3. Summary of Optimal Rearing Conditions for the Various Striped Bass Life Stages Eggs Pro larva Larvae Postlarvae Juveniles & Subadults Adults ABIOTIC FACTORS Temperature Salinity Dissolved Oxygen Light Turbidity BIOTIC FACTORS Diet Density 16-20°C 16-21°C 2-IQO/oo 5-150/oo air saturated natural photoperiod 500 mg/1" not applicable not applicable 50- 75 per liter 50-25 per liter air saturated natural photoperiod 18-22°C >10and<25°C 10-200/00 70-300/oo air saturated natural photoperiod <4 nig/ft 15-20% body weight 5-8% body weight (dry) twice daily (wet) per day 30- 1 0 per liter 1 0-2 bass per 100 liters >10and<24°C 70-300/oo air saturated natural photoperiod 3-5% body weight (wet) daily 2.4 g/l maximum a Fine grained sediment. b Bentonite. 25- 20- 6 OS ID- Humphries and Gumming. 1973 Mansueti, 795. Rhodes and Merriner, 1973 Rogers et al., 1977 10 I 20 30 40 Days after hatching 50 60 70 Figure 2. A comparison of growth rates observed under fixed temperature regimes (Rogers et al.. 1977) with those obtained in earlier studies under conditions of increasing temperature. D. Handling juveniles. 1) Juveniles, if needed for research, can be reared from eggs or lar- vae, or collected by seining in spawning rivers. 2) Juveniles collected from the field should be kept separate from any reared or other col- lected bass already in the cul- ture system. 3) Water quality should be moni- tored daily and maintained at optimum conditions (Table 3). 4) Juveniles can be fed frozen brine shrimp, ground squid, prepared diets, or commercial trout feeds; the first is generally preferred. Conclusions During the course of this study all of the life stages of the striped bass from egg to adult were successfully maintained under laboratory conditions. The temperature, salinity, dissolved oxygen, light and tur- bidity requirements of all life stages were either determined empirically, approxi- mated from environmental data, or where reported by other workers corroborated in our laboratory. Optimum and survival limits for each of these parameters were, where appropriate, specified. By main- taining conditions within these bounds, striped bass eggs were repeatedly reared through to the juvenile stage. A popula- tion of striped bass adults was success- fully maintained in captivity for five years. Despite repeated attempts, we were unable during the course of this study to successfully induce spawning in the laboratory. Sexually mature adults of both sexes, however, did occur among our captive population. A step- by-step culture methodology has been prepared for use by future workers. References Bonn, E.W., W.M. Bailey, J.D. Bayless, K.E. Erickson, and R.E. Stevens(eds.). 1976. Guidelines for striped bass cul- ture. Striped Bass Committee, South- ern Division American Fisheries Society. 103 p. Doroshev, S.I. 1970. Biological features of the eggs, larvae and young of the striped bass (Roccus saxatilis [Wal- baum]) in connection with the problem of its acclimation in the U.S.S.R. J. Ichth. 10(2):235-278. ------- Helf rich, P. 1973. The feasibility of brine shrimp production on Christmas Island. U.S. Sea Grant Technical Report UNIHI- SEA-GRANT-TR-73-02. 1 73 p. Humphries, E.T., and K.B. Gumming. 1973. An evaluation of striped bass fingerling culture. Trans. Amer. Fish. Soc., 102(1):13-20. Laurence, G.C. 1977. Caloric value of some North Atlantic calenoid cope- pods. U.S. Fish Bull., 75:218-220. Mansueti, R. 1958. Eggs, larvae, and young of the striped bass, Roccussax- atilis. Ches. Biol. Labs. Contr. No. 112:1-35. Paffenhofer, G.A. 1967. Caloric content of larvae of the brine shrimp Anemia salina. Helg. Wiss. Meer., 16:130- 135. Pearson, J.C. 1938. The life history of the striped bass, or rockfish, Roccus saxatilis (Walbaum). U.S. Bureau of Fish. Bull., 28(49):825-851. Raymont, J.E., J. Austin, and E. Linford. 1963, Biochemical studies on marine zooplankton. I. The biochemical com- position of Neomys/s integer. J. Cons. Inter. Explor. Mer., 28(1):354-363. Rhodes, W., and J. V. Merriner. 1973. A preliminary report on closed system rearing of striped bass sac fry to fin- gerling size. Prog. Fish Cult., 35(4): 199-201. Rogers, B.A., D.T. Westin, and S.B. Saila. 1977. Life stage duration in Hudson River striped bass. University of Rhode Island Marine Technical Report 31. 11 1 p. Shannon, E.H. 1970. Effect of tempera- ture changes upon developing striped bass eggs and fry. Prog. 23rd Ann. Conf. S.E. Assoc. Game & Fish Comm. pp. 265-274. Slobodkin, L.B., and S. Richman. 1961. Calories/gm in species of animals. Nature (London). 191:299. Table 4. Caloric and Percent Composition of Some Live Larval Food Items Food Item Artemia salina nauplii adults Acartia clausi Acartia tonsa Calanus finmarchicus Calanus helgolandicus Calories/gram (ash-free, dry) 5800-6000(1)* 5454-5953(3) 5115-5854(3) 5664 ± 8612) 6835 ± 191(2) 5515 ± 277(5) Percent of Dry Weight Lipid Protein 15.04-27.24 6.51 5.8 10.5-47.0 11.5 42.5-50. 2(1 J 62.78(1) 82.6(4) 30-77(4) 75.2(4) *Numbers in parentheses refer to source: (1) Helfrich et al. (1973); (2) Laurence (1977); (3) Paffenhofer (1967); (4) Raymont et al. (1963); and (5) Slobodkin and Richman (1961). Bruce A. Rogers, Deborah T. Westin, and Saul B. Saila are with the Graduate School of Oceanography, University of Rhode Island, Kingston, Rl 02881. Allan D. Beck is the EPA Project Officer (see below). The complete report, entitled "Development of Techniques and Methodology for the Laboratory Culture of Striped Bass, Morone saxatilis/' (Order No. PB 82-217 795; Cost: $22.50, subject to change) will be available only from: National Technical Information Service 5285 Port Royal Road Springfield, V'A 22161 Telephone: 703-487-4650 The EPA Project Officer can be contacted at: Environmental Research Laboratory U.S. Environmental Protection Agency South Ferry Road Narragansett. Rl 02882 ------- United States Environmental Protection Agency Center for Environmental Research Information Cincinnati OH 45268 Postage and Fees Paid Environmental Protection Agency EPA 335 Official Business Penalty for Private Use $300 PS 0000329 U S tNVJR FHOTtCTION KtfciON b LldHAHY 230 S UEARbORw STRtET CHICAGO IL 60604 ------- |