ICES mar. Sei. Symp., 198: 626-631. 1994 Survival of eggs and yolk-sac larvae of Baltic cod (Gadus morhua L.) at low levels in different salinities Anders Nissling Nissling, A. 1994. Survival of eggs and yolk-sac larvae of Baltic cod (Gadus morhua L.) at low levels in different salinities. - ICES mar. Sei. Symp., 198: 626-631. In the Baltic Sea, successful spawning of cod is restricted to the deep basins, the Bornholm, Gdansk, and Gotland basins, with salinities of 10-17. Deep water is exchanged mainly during periods of inflow of saline water from the North Sea. These inflows are irregular and stagnant waters prevail for long periods accompanied by unfavourable conditions. Consequently, cod eggs occur regularly at low levels. In this study, batches of eggs (2, 4, and 7 days old) and larvae (newly hatched and 4 days old) were exposed to low concentrations - 1.0-3.5 mg 0 2/lat two salinities - 11 and 15 - and compared with survival in controls at 100% saturation in short-term experiments. Egg survival decreased with decreasing concentration, with no difference in tolerance between egg stages. Egg survival at low levels was clearly affected by salinity, with significantly higher survival at 15 ppt than at 11 ppt. This implies that egg survival at low levels differs between the spawning grounds of Baltic cod. It is higher in the Bornholm Basin with higher salinities, 13-17, than in the Gdansk and Gotland basins, with 10-13. For both larval stages, a marked decrease in tolerance to low levels was evident between 3.0 and 2.5 mg 0 2/l irrespective of salinity, suggesting that the limit for survival of Baltic cod larvae is at about 2.5-3.0 mg 0 2/l. Anders Nissling: Department o f Systems Ecology, Stockholm University, S-106 91 Stockholm, Sweden. Introduction Successful spawning of Baltic cod is restricted to the Baltic deep basins - the Bornholm, Gdansk, and Gotland basins. In the Baltic deep basins there is a halocline at 50-70 m depth with a denser more saline (10-17 ppt) deep water which is only partly mixed with the less saline (6-8 ppt) surface water. The deep water is exchanged mainly during occasions of inflow of saline water from the North Sea. These inflows are irregular, however, and during periods between inflows stagnant conditions prevail with a decrease in the salinity and content of the water. These varying conditions have a profound impact on the reproductive success of fishes with pelagic eggs. In the Baltic, cod eggs often occur in layers with low content (see Lebedek, 1978; Wieland, 1989), as determined by their buoyancy. The year-class strength of Baltic cod is thought to be established during the embryonic period and apparently determined by the thickness of the spawning layer (Grauman, 1973), i.e., the layer with 11 ppt salinity or more (Westin and Nissling, 1991) and sufficient content for egg and larval development. The last two major inflows of saline water to the Baltic Sea occurred in the winters 1975-1976 and 1976-1977 (Fonselius, 1988) and resulted in favourable spawning conditions. Consequently, an increase in the Baltic cod stock followed and high catches were reported from the late 1970s until the mid-1980s [450 000 t in the early 1980s (Hansson and Rudstam, 1990)]. However, since stagnant water has prevailed in the deep basins from the late 1970s, there has been a dramatic decrease in the Baltic cod stock, with poor catches reported since the mid-1980s [about 75000 t reported in 1992 (P.-O. Larsson. pers. comm.)]. The metabolic rate of fish embryos is independent of the water content until a certain critical level is reached, below which metabolism becomes dependent on ambient concentrations (Rombough, 1986). Any deviation from normal uptake will influence development and survival negatively (Serigstad, 1987). Metabolic rate in fishes is highly influenced by temperature, since temperature controls the rate of different biochemical reactions, and by salinity, which affects osmoregulation. Consequently, requirements of eggs and larvae are affected by temperature [see Schur-
ICES mar. Sei. Symp.. 198(1994) Survival o f eggs and yolk-sac larvae o f Baltic cod 627 mann and Steffensen, 1992 (juvenile cod); Alderdice and Forrester, 1971 (Pacific cod eggs); Almatar, 1984 (herring and plaice larvae)] and possibly by salinity (see Alderdice and Forrester, 1971; Almatar, 1984). The aim of the present investigation was to study the tolerance of Baltic cod eggs and larvae to low concentrations at prevailing salinities of the Baltic deep basins. In short-term experiments (48 h), eggs (three different stages) and yolk-sac larvae (two stages) were incubated at low levels; 1.0-2.8 (for eggs) and 1.5-3.5 (for larvae) mg 0 2/l at 7 C at 11 and 15 ppt and compared with survival in controls at 100% saturation. 1a. C 60 control 15 ppt 0 11 ppt Materials and methods Spawning cod were caught in gillnets at 50-90 m depth off Gotland (58 N19 E) in the middle of the Baltic Sea. Eggs and semen were obtained by stripping. Fertilization was carried out in water at 7 C and 17 ppt prepared from filtered (0.2 /im cartridge filter) sea water (7 ppt) and synthetic sea salt (hw Marinemix). Eggs at three different stages of development (2, 4, and 7 days old) and larvae at two stages [newly hatched and 4 days old (onset of first feeding)] were incubated. Water of different concentrations was obtained by bubbling with nitrogen gas to the nearest 0.1 mg 0 2/l (WTW Oximeter 196). In all incubations, filtered water was treated with antibiotics [Mycostatin (2500 IU/1), Streptomycin (0.05 g/1), and Doctacillin (0.2 g/1)]. Treated water was transferred to the incubation flasks by means of a tube before the eggs and larvae were added. Prior to incubation the eggs and larvae were kept in water at 7 C and 17 ppt salinity. Batches of eggs and larvae were incubated at five concentrations, ranging between 1.0 and 2.8 mg 0 2/l for eggs and 1.5 and 3.5 mg 0 2/l for larvae, at two different salinities; 11 and 15 at 7 C. As controls, incubations at 100% saturation (10-11 mg 0 2/l) in respective salinities were used. Each treatment, containing 125-150 eggs or 10-15 larvae, was incubated for 48 h in a 500 ml flask with conical stopper to prevent air bubbles at the start of incubation. The water was mixed gently three times a day by rotating the flask to prevent microstratification of. Each set of incubation, i.e., 12 flasks, was performed in replicates with batches from different females as follows: 6 replicates for 2 and 4 days old eggs; 2 replicates for 7 days old eggs; 5 and 6 replicates for newly hatched and 4 days old larvae, respectively. The flasks were placed on their side and the eggs developed at the bottom of 11 ppt, since Baltic cod eggs are non-buoyant at 11 ppt (see Nissling and Westin, 1991); because of positive buoyancy they developed at the top at 15 ppt. At the end of the incubations egg 1b. 100 80 1c. 100 60 TT 0 11 ppt control / / 2,4 2,0 1,7 1,4 1,0 1 control! / 2,4 2,0 1,7 1.4 B 15 ppt 0 11 ppt Figure 1. Survival of Baltic cod eggs incubated at low levels for 48 h at 7 C with salinities of 11 and 15. Survival in controls is given as absolute survival and survival at low concentrations is given as relative survival, i.e. as percentage of survival in controls, (a) 2 days old eggs (six replicates), (b) 4 days old eggs (six rcplicates), (c) 7 days old eggs (two replicates). Standard deviation shown at top of bars.
628 A. Nissling ICES mar. Sei. Symp., 198 (1994) Table 1. Oxygen concentrations (mg 0 2/l) at the start and end of exposure of Baltic cod eggs (2 and 4 days old) (six replicates) and larvae (4 days old) (one larval group) to low levels. Each experiment was carried out at 7 C with a salinity of 11 for 48 h. Eggs Larvae Start of incubation (mg (VI) End of incubation (mg 0 2/l) 2 days old 4 days old Start of incubation (mg 0 2/l) End of incubation (m g 0 2/l) 2 days old 10-11 (control) 9.0 ± 0.6 9.3 ± 0.4 10-11 (control) 9.6 2.8 2.6 ± 0.1-3.5 3.4 2.4 2.3 ± 0.1 2.3 ± 0.1 3.0 3.0 2.0 2.0 ± 0.1 1.9 ± 0.1 2.5 2.5 1.7 1.6 ± 0.1 1.6 ± 0.1 2.0 2.0 1.4 1.4 ± 0 1.4 ± 0 1.5 1.5 1.0-1.1 ± 0.1 - survival and larval condition were checked. White eggs (dimpled) and eggs which had ceased in development were considered dead and larval condition was judged as follows: dead, moribund (partly white), alive but inactive (did not react when touched), or active (swimming). In order to exclude differences in survival due to differences in egg quality caused by maternal effects and stage during the spawning period (see Kjesbu, 1989; Kjesbu et al., 1991; Solemdal et al., 1991), egg survival was calculated in relation to survival in the controls (survival in the controls treated as 100% survival) and given as a percentage of survival in the controls. To check the consumption of during the incubation, concentrations at the end of incubations at 11 ppt were measured. To examine whether transference from a high salinity (17) to lower salinities (11 and 15) influenced egg survival at low concentrations, two egg batches were fertilized and incubated at respective salinity until start of exposure, i.e., day 3 (one batch) and day 5 (one batch), and then incubated at low concentrations at 7 C for 48 h as above. Results Table 1 gives the concentrations at the end of incubations for 2 and 4-day-old eggs and one larval group. The measurements revealed a small decrease in content during the 48 h incubation. The highest decrease occurred in the controls, while the decrease was low at low levels. The decreases in content seem to be related to egg survival, i.e., the higher the survival the higher the consumption (cp. with Fig. 1). Egg survival and larval condition after incubation at low concentrations at 11 and 15 ppt are shown in Figures la-c and 2a-d, respectively. Egg survival decreased with decreasing concentrations at all developmental stages studied, with no apparent difference in tolerance to low levels between egg stages. Compared with controls (100% saturation) there was a decrease in egg survival at 2.8 mg O2/ 1and only a few eggs survived at 1.0 mg 0 2/l. However, egg survival differed between salinities and was significantly higher at 15 ppt than at 11 ppt, p < 0.001 (sign test of paired observations) for both 2 and 4-day-old eggs (the numbers of incubations of 7-day-old eggs were too low for a sign test) at low concentrations, while no difference in survival occurred between controls at 11 and 15 ppt. Table 2 gives egg survival at low concentrations at 11 and 15 ppt for two egg batches incubated at the same salinities from fertilization until the start of exposure to low concentrations. Egg survival was higher at 15 ppt than at 11 ppt for both 3 and 5-dayold eggs. These results suggest that the difference in egg survival between salinities was not caused by the transference of eggs from a higher salinity (17 ppt) at the start of exposure. In short-term incubations of larvae at low concentrations at 11 and 15 ppt, no differences in tolerance to low concentrations were apparent be- Table 2. Survival of Baltic cod eggs (3 and 5 days old) fertilized and incubated with salinities of 11 and 15 and exposed to low levels for 48 h at 7 C at identical salinities. Oxygen (mg 0 2/l) 3 days old 5 days old 11 ppt 15 ppt 11 ppt 15 ppt 10-11 (control) 96.5 95.5 73.6 70.4 2.4 34.5 67.8 22.5 58.8 2.0 16.1 36.1 5.0 20.7 1.7 3.5 31.8 4.1 9.4 1.4 0-1.1 7.5 1.0 0 7.2 - -
ICES mar. Sei. Symp., 198 (1994) Survival of eggs and yolk-sac larvae o f Baltic cod 629 alive active E3 alive inactive H moribund I dead control 3,5 3,0 2,5 2,0 1,5 control 3,5 3,0 2,5 2,0 1,5 Fig. 2c. Fig. 2d. 100 90 80 70-60- 50-40- 30- D alive active alive inactive 9 moribund dead 20 1 0 - control 3,5 3,0 2,5 2,0 0 control 3,5 3,0 Figure 2. Condition of Baltic cod larvae incubatcd at low levels for 48 h at 7 C. (a) Newly hatched larvae incubated with a salinity of 11 (five replicates), (b) newly hatched larvae incubated with a salinity of 15 (five replicates), (c) 4 days old larvae incubated with a salinity of 11 (six replicates), (d) 4 days old larvae incubated with a salinity of 15 (six replicates). tween larval stages or between incubation salinities. For both newly hatched larvae and larvae at the onset of feeding the majority of larvae were alive and active at 3.5 mg 0 2/l, with inactive and moribund larvae beginning to appear at 3.0 mg 0 2/l. For both larval stages there was a marked decrease in tolerance to low concentrations between 3.0 and 2.5 mg 0 2/l. Incubations at 3.0 mg 0 2/l displayed high survival, whereas incubations at 2.5 mg 0 2/l or lower revealed dead or moribund larvae with only a few surviving larvae at 2.5 mg 0 2/l. Discussion Surveys in the Baltic deep basins have shown that adult cod may occur in layers with content as low as 1.4-2.9 mg 0 2/l (see Berner and Schemainda, 1957, 1958; Tiews, 1970) and that cod eggs are regularly found at low levels; 1.7-5.4 mg 0 2/l (Lebedek, 1978), 2.6-5.4 mg 0 2/l (Wieland, 1989) with high mortalities reported during the egg stage period; 79-98.8% (Grauman, 1973), 99.8% (Wieland, 1987). The critical
630 A. Nissling ICES mar. Sri. Symp., 198 (1994) level for successful development of Baltic cod eggs has been suggested as being 1.4 mg 0 2/l (Grauman, 1973); >2.9 mg 0 2/l (Kosior and Netzel, 1989), but experimental data are scarce. (Oxygen levels reported in ml 0 2/l are recalculated to mg 0 2/l). The present study shows that cod eggs may survive at low levels for at least two days, but that survival is greatly affected at concentrations of 1.0-2.8 mg 0 2/l. Since cod eggs are regularly found at levels of this magnitude, these results suggest that low levels in the Baltic deep basins may cause high egg mortalities. As concluded by Wieland, from investigations on cod egg distribution in the Bornholm Basin, vertical occurrence of eggs is not only controlled by egg buoyancy. In addition to water density, the vertical occurrence of eggs is also influenced by levels as a result of high egg mortality at low concentrations. Cod eggs were most abundant at depths of 60-75 m at levels of 2.6-6.9 mg 0 2/l, but eggs also occurred deeper, at concentrations of less than 1.4 mg 0 2/l (Wieland. 1987, 1989). According to several investigations on marine fish embryos the uptake rate of increases during development from fertilization to hatching and absorption of the yolk sac [Serigstad, 1987 (cod); Davenport and Lönning, 1980 (cod); Finn etal., 1991 (halibut)], and is also influenced by larval activity level (Davenport and Lönning, 1980; Serigstad, 1987), i.e., an increase of demand during development. However, in this study no differences in tolerance to low levels among egg stages or between two larval stages were observed. For both larval stages a marked decrease in tolerance to low concentrations was evident between 3.0 and 2.5 mg 0 2/l, with only a few surviving larvae at 2.5 mg 0 2/l. Accordingly, this suggests that the limit for survival of Baltic cod larvae is at about 2.5-3.0 mg 0 2/l. Incubation of eggs and larvae at two different salinities showed that larval condition was not affected by salinity at low levels, whereas egg survival at low concentrations was significantly higher at 15 ppt than at 11 ppt. Hence, the higher mortality of eggs at low levels at 11 ppt compared to 15 ppt, whereas no differences in egg mortality occurred between controls (100% saturation), suggests that cod eggs are more vulnerable to deficit at extreme salinities. Further, this investigation indicates that Baltic cod are less affected by extremes in salinity as larvae than at the egg stage. Although a long-term experiment is required to estimate overall egg mortality from fertilization to hatching at a certain level, the present investigation indicates a significant difference in total egg mortality between 11 and 15 ppt. For instance, if egg mortality at 48 h of exposure at 2.4 mg 0 2/l (egg mortality at the three egg stages pooled for 11 and 15 ppt) is extrapolated, it results in an overall mortality (egg development time 13-15 days at 7 C) of about >99.99 and 97% at 11 and 15 ppt, respectively. This indicates a considerable difference in spawning success with salinity for a given concentration. The inflows of saline rich water from the North Sea are irregular and as a consequence conditions for successful spawning vary greatly between years, but also between spawning areas. Because of the relative distance from the North Sea, the most favourable conditions, generally higher concentrations and more saline water (13-17 ppt compared with 10-13 ppt in the Gdansk and Gotland basins), occur in the Bornholm Basin, which is more regularly influenced by influxes of water from the North Sea than are the Gdansk and Gotland basins, where stagnant periods are more pronounced. Consequently, egg survival is higher in the Bornholm Basin than in the Gdansk and Gotland basins, owing to more favourable conditions but also, as this study implies, because of higher salinity, since eggs are less vulnerable to depletion at 15 ppt than at 11 ppt. Acknowledgments The investigation was financially supported by the Swedish Council for Forestry and Agricultural Research and the National Swedish Board of Fisheries. References Aldcrdice, D. F., and Forrester, C. R. 1971. Effects of salinity, temperature, and dissolved on early development of Pacific cod (Gadus macrocephalus). Fish. Res. Bd Can., 28: 883-902. Almatar, S. M. 1984. Effects of acute changes in temperature and salinity on the uptake of larvae of herring (Clupea harengus) and plaice (Pleuronectes platessa). Mar. Biol., 80: 117-124. Berner, M., and Schemainda, R. 1957. Über den Einfluss der hydrographischen Situation insbesondere des Durchlüftungszustandes - auf die vertikale Verteilung und den Fang der Laichdorschschwärme im Bornholmbecken. Z. Fisch. N.F., 6: 331-342. Berner, M., and Schemainda, R. 1958. Über die Abhängigkeit der Laich dorscherträge im Bornholmbecken von der hydrographischen Situation. Dt. Fischerei Z., 5(3): 65-70. Davenport, J., and Lönning, S. 1980. Oxygen uptake in developing eggs and larvae of the cod, Gadus morhua L. J. Fish Biol., 16: 249-256. Finn, R. N., Fyhn, H. 1., and Evjen, M. S. 1991. Respiration and nitrogen metabolism of Atlantic halibut eggs (Hippoglossus hippoglossus). Mar. Biol., 108: 11-19. Fonselius, S. 1988. Long-term trends of dissolved, ph and alkalinity in the Baltic deep basins. ICES CM 1988/C:23, pp. 1-6. Grauman, G. B. 1973. Investigations of factors influencing fluctuations in the abundance of Baltic cod. Rapp. P.-v. Réun. Cons. int. Explor. Mer, 164: 73-76.
ICES mar. Sei. Symp., 198 (1994) Survival o f eggs and yolk-sac larvae o f Baltic cod 631 Hansson, S., and Rudstam, L. G. 1990. Eutrophication and Baltic fish communities. Ambio, 19: 123-125. Kjesbu, O. S. 1989. The spawning activity of cod, Gadus morhua L. J. Fish Biol., 34: 195-206. Kjesbu, O. S., Klungsøyr. J., Kryvi, H.. Witthames. P R., and Greer Walker, M. 1991. Fecundity, artesia, and egg size of captive Atlantic cod (Gadus morhua L.) in relation to proximate body composition. Can. J. Fish, aquat. Sei., 48: 2333-2343. Kosior, M.. and Nctzcl. J. 1989. Eastern Baltic cod stocks and environmental conditions. Rapp. P.-v. Réun. Cons. int. Explor. Mer, 190:159-162. Lebedek, W. 1978. Occurrence and distribution of sprat and cod eggs and larvae in the southern Baltic in 1977. ICES CM/ J:15, pp. 1-4. Nissling, A., and Westin, L. 1991. Egg buoyancy of Baltic cod (Gadus morhua) and its implications for cod stock fluctuations in the Baltic. Mar. Biol., Ill: 33-35. Rombough, P. J. 1986. Mathematical model for predicting the dissolved requirements of steelhead (Salmo gairdneri) embryos and alevins in hatchery incubators. Aquaculture, 59: 119-137. Schurmann, H., and Steffensen, J. F. 1992. Lethal levels at different temperatures and the preferred temperature during hypoxia of the Atlantic cod, Gadus morhua L. J. Fish. Biol., 41: 927-934. Serigstad, B. 1987. Oxygen uptake of developing fish eggs and larvae. Sarsia, 72: 369-371. Solemdal, P., Kjesbu, O. S., Opstad, I., Skiftesvik, A. B., Birkeland, R., Bratland, P., and Fonn, M. 1991. A comparison of egg quality and larval viability between eultured coastal cod and wild Arcto-Norwegian cod. ICES CM/F:41, pp. 1-13. Tiews, K. 1970. Über die Verbreitung des Laichdorschbestandes in der mittleren Ostsee in Abhängigkeit vom Sauerstoffgehalt in den Jahren 1962-1970. Arch. Fischereiwiss., 21:213-221. Westin, L., and Nissling, A. 1991. Effects of salinity on spermatozoa motility, percentage of fertilized eggs and egg development of Baltic cod (Gadus m orhua), and implications for cod stock fluctuations in the Baltic. Mar. Biol., 108: 5-9. Wieland, K. 1987. Distribution and mortality of cod eggs in the Bornholm Basin (Baltic Sea). ICES CM/G:56, pp. 1-14. Wieland, K. 1989. Small-scale vertical distribution and mortality of cod and sprat eggs in the Bornholm Basin (Baltic Sea) during two patch studies in 1988. ICES CM/J:19, pp. 1-15.