Induction of trypsinogen secretion in herring larvae (Clupea harengus)

Mechanisms initiating trypsinogen secretion were studied in laboratory reared herring larvae (Clupea harengus L.) exposed to physical and chemical stimuli. Pancreatic secretion of trypsinogen was quantified for each stimulus type as the increase above pre-stimulus level of intestinal trypsin content. Larval prey types were: nauplii, copepodites or adult Acartia tonsa, small polystyrene spheres (diameter 94 m), small (diameter 79 m) or large (diameter 170 m) polystyrene-latex spheres. Intestinal trypsin content can be expressed as a function of two variables: meal size and content of pancreatic trypsinogen. Trypsinogen secretion increases with different prey items in the order: small spheres, nauplii and copepodites. Larvae which eat large spheres secrete more enzyme than if fed small spheres but trypsinogen secretion is similar in fish larvae fed copepodites and large spheres. The fact that the size of non-biodegradable particles exerts a major control over trypsinogen secretion suggests neural — as opposed to chemically mediated — initiation of secretion. A cephalic phase of secretory stimulation could not be demonstrated during swallowing of copepods or exposure for 2 to 3 h to compounds which leak from live copepodites. As cephalic and gastric phases of secretory stimulation are absent, initiation of trypsinogen secretion must take place in the intestine. Larval herring retain trypsin in the intestine. Ca. 4.5 h after a meal, 3/4 of the enzyme is located in the intestinal fluid, presumably available for hydrolysis of subsequent meals, and the high proportion (ca. 25%) of the pancreatic trypsinogen content which is secreted for copepodite prey may thus not be energetically wasteful for the larvae.     

Developmental changes in the composition of myofibrillar proteins in the swimming muscles of Atlantic herring,Clupea harengus

Changes in myofibrillar protein composition during development have been investigated in the swimming muscles of the Atlantic herring Clupea harengus L. using a range of electrophoretic techniques. The main muscle-fibre type of larvae, and the fast- and slow-muscle fibres of adult fish were found to contain distinct isoforms of myosin heavy chain (MHC) and myosin light chain 2 (LC2). Larval LC2 was present as a minor component of adult fast-muscle myosin. In contrast, larval and adult fast-muscle myosin appeared to contain identical alkali light chains. Tropomyosin and troponin C were also identical in larval and in adult fast-muscle. All three muscle-fibre types contained unique isoforms of troponin T (TNT) and troponin I (TNI). Larval muscle had multiple isoforms of TNT, some of which may correspond to embryonic forms. It was concluded that although the main muscle-fibre type in larvae shares some myofibrillar proteins with adult fast muscle, it also contains characteristic isoforms of MHC, TNI, TNT and LC2 and therefore represents a distinct fibre type. The particular combination of myofibrillar proteins present at any developmental stage was found to be dependent on the rearing temperature. For example, a higher proportion of embryonic TNT isoforms were present at hatching in larvae reared at 5°C than at either 10 or 15°C. Over a period of 7 d, there was a gradual reduction in the number of TNT isoforms, but the pattern in 5°C larvae after 7 d still did not resemble that in 1 d-old larvae reared at 15°C.     

Feeding ecology of herring (Clupea harengus) larvae in the turbid Blackwater Estuary

Most studies on feeding by herring larvae (Clupea harengus) have taken place in clear, open waters, but several herring stocks around the world spawn in inshore and estuarine regions. An example is the spring-spawning Blackwater Estuary (Essex, England) stock. Samples were collected in this estuary to examine prey selectivity and feeding levels in relation to biological and environmental conditions. Herring larvae negatively selected copepod nauplii, but positively selected the copepodite and adult stages of Acartia spp. Gastropod larvae were also positively selected. Particles >150 μm width were preferred, whilst particles smaller than this value were preferentially rejected. Concentrations of potential prey items in the water were in the range of 6.0 to 49.7 organisms l⁻¹ with a median concentration of 15.0 organisms l⁻¹ (n = 26). These values are towards the low end of prey concentrations quoted in the literature as being required to sustain herring larval growth and survival. However, theoretical considerations suggest that, in this environment, levels of tidally-induced turbulence enhance encounter rates between larval herring and their prey. On the other hand, turbidity is also related to tidal current speed and might reduce feeding success by decreasing underwater light levels. Measurements at two sites in the estuary confirmed that tidally-induced turbidity reduced the effective water depth in which herring larvae could visually feed by up to 50% at times of peak current speed. However, with the gut-content data available in the present study, it was not possible to discern any clear relationships between feeding success and the state of the tide. Feeding success appeared to be more strongly influenced by surface light-levels. Received: 24 June 1998 / Accepted: 17 February 1999     

RNA:DNA ratios and growth of herring (Clupea harengus) larvae reared in mesocosms

Autumn-spawned North Sea herring larvae (Clupea harengus L.) were released in two outdoor mesocosms of 2500 m³ (A) and 4000 m³ (B). The mesocosms were monitored for temperature, salinity, oxygen, chlorophyll a, zooplankton and herring larvae abundance. The density of suitable prey for first feeding larvae (mainly copepod nauplii) was initially low in Mesocosm A (<0.11^-1) compared to in Mesocosm B (>11^-1). Half-way through the experiment the situation was reversed, with higher densities of prey in Mesocosm A (>31^-1) as compared to Mesocosm B (～11^-1). The average temperature declined steadily in both mesocosms from 18°C at release to 11–12°C by the end of the experiment 60 d later. The RNA:DNA values of individual herring larvae were related to protein growth rates and temperature adjusted according to Buckley (1984). A corresponding DNA growth index (Gdi) was given as: Gdi=0.68 TEMP+3.05 RNA:DNA-9.92. The RNA:DNA based growth indices were significantly correlated with other somatic growth estimates. The average estimated protein growth rate in the two mesocosms followed the same temporal pattern as the somatic growth rate, but with a lag of 2 d or more. Residual analysis of the regression of ln RNA versus ln DNA also showed the same temporal pattern as the RNA:DNA ratios, but the shift in condition as estimated by this method occurred more in synchrony with the other somatic growth measures. Larvae in Mesocosm A had RNA:DNA values similar to the starvation control kept in the laboratory the first days after release, confirming that larvae in Mesocosm A initially were in poor nutritional condition. On the other hand, the majority of the herring from Mesocosm B were characterised as starving or in poor nutritional condition towards the end of the experiment. The assessment of growth and nutritional condition were in accordance with independent survival estimates which suggested that the majority of the total mortality occurred during the first 15 d in Mesocosm A and there-after in Mesocosm B.     

High natural mortality of Baltic herring (Clupea harengus) eggs caused by algal exudates?

Results from field studies in 1978 and 1982 together with results from a laboratory study carried out in 1984 indicate that the unusually high natural mortality of Baltic herring (Clupea harengus L.) eggs observed in situ is presumably caused by toxic exudates released by filamentous brown algae during a limited period which happens to coincide with the peak of spawning. Oxygen deficiency is no longer believed to be the major cause of the observed unnaturally high mortality. It cannot, however, be entirely ruled out.     

Escape behaviour of solitary herring (Clupea harengus ) and comparisons with schooling individuals

The escape behaviour of solitary herring (Clupea harengus L.) startled by a sound stimulus was observed by means of high-speed video-filming. The results were compared with data from a previous study on the escape behaviour of schooling herring. Escape responses were divided into “away responses” and “towards responses” according to the orientation of the C-bend of the body relative to the stimulus. The proportion of away responses was smaller for solitary than for schooling herring. In solitary herring, the subsequent escape trajectories of fish making initial away responses showed a bimodal pattern of distribution, with modes at 130 and 180° from the stimulus. Trajectories following towards responses, however, were mainly within the semicircle directed at the stimulus, and their pattern of distribution differed from that of away responses. This result contrasts with observations on schooling herring, whose trajectories following both initial away and towards responses are directed away from the stimulus. In addition, we measured the response latency, defined as the interval of time between stimulus presentation and the first detectable movement of the fish. Solitary herring showed a higher proportion of short-latency responses (latency <50 ms) than schooling herring. Different behaviours appear to be exhibited by herring depending on whether they are solitary or within a school. We hypothesize that schooling may raise the threshold for initiation of fast escape responses, giving longer latencies and slower responses which are more appropriate in their directionality and reduce the possibility of collisions with neighbours. In addition, we suggest that schooling behaviour enhances the directionality and co-ordination of the escape response of the whole school, possibly increasing the probability of surviving a predator attack. Received: 2 October 1996 / Accepted: 17 October 1996     

Temperature influences muscle differentiation and the relative timing of organogenesis in herring (Clupea harengus) larvae

Eggs from spring spawning stocks of herring (Clupea harengus L.) were fertilized and reared at either 5, 8 or 12°C in 1991 and 1992. The differentiation of myotomal muscle fibres was investigated in relation to the development of other organs and tissues using light and electron microscopy. The gut, notochord, eyes and haemocoel appeared at the same relative point in development between fertilization and hatching at all temperatures. In contrast, the formation of the spinal cord, pronephros, pectoral fin buds and muscle fibres was relatively retarded at 5°C compared with 8 and 12°C. Myogenesis in the presumptive inner muscle mass occurred after 12 to 16 d at 5°C, 7 to 10 d at 8°C and 3.5 to 6 d at 12°C. Myoblasts aligned in orderly rows running from myosept to myosept prior to fusion to form myotubes. Actin and myosin filaments were synthesised throughout the cytoplasm in associated with presumptive Z-lines at the periphery of myotubes and immature muscle fibres. Differentiation of the superficial and inner muscle fibres types of larvae occurred at around the same time. Following this initial period of myogenesis, the number of myotomal muscle fibres remained constant until after hatching, so that increases in muscle bulk in the late embryo were entirely due to fibre hypertrophy. At hatching, the number of superficial muscle fibres present in myotomes just posterior to the yolk-sac was significantly less at 5°C (108±12) than at either 8°C (132±10) or 12°C (140±10) (mean±SD, 12 fish/temperature). In contrast, there were around 280 inner muscle fibres/myotome, comprising 90% of the trunk cross-sectional area, at all three temperatures. Myofibrillargenesis occurred relatively slowly at low temperatures, so that the volume density of myofibrils in the inner muscle fibres of larvae at hatching was significantly less at 5°C (39.2±9.0) than at either 8°C (49.6±8.8) or 12°C (50.2±9.8) (mean ±SD, 20 fibres/temperature from total of 5 fish). Undifferentiated myoblasts remained at hatching to form a population of presumptive myosatellite cells. The number of presumptive myosatellite cells per mm² cross-sectional area of muscle fibre was more than two times higher at 8°C (1493±335) than at either 5°C (478±102) or 12°C (924±233) (mean±SD, 5 fish/temperature). The results suggest that temperature can influence the commitment of myoblasts to differentiation at a critical stage in embryogenesis, thereby providing a potential mechanism for influencing future growth characteristics. Correspondence to: I.A. Johnston at Gatty Marine Laboratory     

Variations in the content of trypsin and trysinogen in larval herring (Clupea harengus) digesting copepod nauplii

Trypsin and its proform trypsinogen were quantified by radioimmunoassay in herring (Clupea harengus L.) larvae subjected to different prey densities. During the first weeks of larval life, the enzyme content fluctuated in a threephased pattern. Yolk resorption (Phase 1) was characterized by an increase in enzyme. During the first few days after yolk resorption (Phase 2), there was a sharp decline in enzyme. Older larvae (Phase 3) exhibited a second period of intensive enzyme synthesis. Amounts of trypsin in intestines of feeding larvae were analysed. At first feeding, a basal level of gut enzyme of approximately 30ng was recorded, and the amount of additional enzyme secreted from the pancreatic tissue into the intestine appeared to be dependent upon the numbers of prey items ingested. The enzyme-substrate ratio in the intestine was approximately 1 to 4. Prey availability affected amount of trypsinogen. Larvae experiencing a high prey density had an approximately two-fold higher specific enzyme content in Phase 2 compared to larvae exposed to a low prey density. A proposed nutritional strategy for first feeding herring larvae is discussed.     

Effects of sulfuric water pollutants on fertilization,embryonic development and larvae of the herring,Clupea harengus

Using herring (Clupea harengus) eggs and larvae from Baltic spring spawners, the biological effects of sulfuric pollutants (largely FeSO₄ and H₂SO₄) which are scheduled to be released daily into the North Sea in large quantities, have been tested. Dilutions (1:8000, 1:16000, 1:24000 and 1:32000) of the sulfuric pollutants were used as test media. Throughout all experiments water temperature was maintained at 8.0°±0.2°C, salinity near 16.5; eggs were artificially fertilized 1 or 6 to 7 h after catching the parent individuals and, attached to glass plates, incubated in 1 l aerated containers. In all test media brownish precipitates, which resulted from diluting the sulfuric pollutants with 50% sea water, tend to adhere to the egg surfaces, thereby possibly interfering with gaseous and other exchanges between egg and surrounding medium. Under conditions of maximum test medium effectiveness, percentages of successful fertilization and of egg survival are considerably reduced; diameter of fertilized eggs remains smaller; embryonic growth rate is retarded while the heart frequency tends to increase (indicating physiological stress); duration of incubation is shortened: percentage of successful hatching decreases and structural abnormalities of freshly hatched larvae increases. Dilutions down to 1:24000 and 1:32000 have rather limited effects. Exposure of 1 to 3 day old, healthy herring larvae to the 4 test media leads to failure of performing prey catching manoeuvres in 1:32000 and 1:24000, to impaired locomotory performances in 1:16000, and practically to paralysis, shrinkage, permanently bent bodies and death within a few days in 1:8000. Although the present study needs deepening through further and more detailed experiments, it can be said that the pollutants under consideration represent a danger to herring eggs and larvae at least up to a dilution of 1:32000.     

Assimilation of the wax esters of marine zooplankton by herring (Clupea harengus) and rainbow trout (Salmo gairdnerii)

Herring (Clupea harengus L.) and rainbow trout (Salmo gairdnerii) fed frozen-thawed zooplankton for 7 days assimilated more than 90% of the dry matter in the zooplankton consumed. Less than 5% of the total lipid ingested, largely wax esters, was excreted in the faeces. Faecal lipids from both the herring and the trout, especially the latter, were rich in wax esters and free fatty alcohols. Faecal fatty alcohols were enriched in 22:1 and deficient in 16:0 and 16:1 alcohols compared to zooplankton fatty alcohols. The growth of rainbow trout fed frozen-thawed zooplankton over 3 months was about one-half of that of trout fed commercial pellets. The fish fed on zooplankton appeared normal and were more pigmented than the fish fed on commercial pellets.     

Otolith ring deposition in relation to growth rate in herring (Clupea harengus) and turbot (Scophthalmus maximus) larvae

Larvae of Clyde spring-spawning Clupea harengus L. and hatchery-produced Scophthalmus maximus (L.) were reared from hatching through metamorphosis in 1980 and 1981 in laboratory tanks and in large enclosures under various light, temperature, and feeding regimes in order to study otolith ring deposition and growth under different conditions. Ring deposition and growth rates were significantly affected by rearing conditions in both species. The ring deposition rates observed under the conditions tested ranged from 0.34 to 0.92 rings d^-1 in herring larvae, and from 0.07 to 1.0 rings d^-1 in turbot larvae. Growth rates ranged from 0.11 to 0.42 mm d^-1 in herring and from 0.05 to 0.27 mm d^-1 in turbot. The number of otolith rings was dependent on the growth rate of the individual larva. At the population level, higher ring deposition rates were observed in faster growing populations. In herring larvae, the relationship between average growth rate and average ring deposition rate was logarthmic, reaching an asymptote at 1 ring d^-1 for growth rates approaching 0.40 mm d^-1. The relationship was linear for turbot larvae for the range of growth rates observed.     

Factors influencing predation of Hyperoche medusarum (Hyperiida: Amphipoda) on larvae of the Pacific herring Clupea harengus pallasi

Predation of different-sized Hyperoche medusarum (Hyperiida: Amphipoda) on larvae of the Pacific herring Clupea harengus pallasi was studied in the laboratory. The attacking rate of H. medusarum was a function of herring larvae size as well as size of the predator, and varied from 0.15 to 0.95 larvae attacked h^-1 per hyperiid. In the range of 7.55 to 16.05 mm total larval length, vulnerability to predation was highest for 13.3 and 13.7 mm larvae. Large hyperiids swam faster and covered a wider area during searching and were more effective predators than small ones. Predation seemed to be influenced by light, and its intensity was dependent on the duration of previous food deprivation of the hyperiid.This study was sponsored by the International Bureau of the Gesellschaft für Kernenergiever-wertung in Schiffbau und Schiffahrt in connection with the German Canadian agreement on scientific and technical cooperation.     

Growth and mortality in young larval herring (Clupea harengus); effects of repetitive changes in food availability

Following yolk resorption, laboratory-reared larval Baltic herring (Clupea harengus L.) were exposed to two sequences of food restriction for 5 d and re-alimentation for 10 d. Comparisons regarding larval growth (standard length and content of water-soluble protein), mortality and content of the sum of trypsin and trypsinogen were made with larvae at a continuous high ration. Larvae exposed to varying prey abundance grew less in length than the control, and during the second high-ration period (Day 22 to 32) growth in length ceased. From the first low-ration period onwards, the content of water-soluble protein in these larvae was lower than that of the control larvae, and the survival rate of the low-high ration group was 59% compared to 77% in the larvae at a continuous high ration. In contrast, the effects of varying food availability were minor on larval content of trypsin and trypsinogen. Results are compared with previous findings in larval Clyde herring, and the effects of larval stock and timing and duration of food restriction on larval growth performance are discussed.     

Cercopagis pengoi and Mysis spp. alter their feeding rate and prey selection under predation risk of herring (Clupea harengus membras)

The anti-predator behaviour of Baltic crustacean planktivores was studied in feeding experiments under predation pressure of herring. The experiments were conducted with pelagic mysids: Mysis mixta and Mysis relicta, and with Cercopagis pengoi, a non-indigenous cladoceran, which invaded the Baltic Sea in 1992. Zooplankton was offered as prey. Two kinds of experiments were performed in the absence and presence of chemical predator cues: (1) two-prey experiments with prey, which have poor or good escape responses and all three planktivores and (2) natural prey experiments with mysids in natural zooplankton assemblages. The results showed that all three species reacted to the chemical cue of herring by decreasing their feeding rate and altering prey selection. C. pengoi selected easily captured prey (rotifers) in two-prey experiments under predation risk while selection for any prey was evident in mysids in natural prey experiments only in the absence of predator cues. This indicates that planktivores have different anti-predator strategies, which are modified by their own prey capture abilities. C. pengoi was a very efficient predator on small prey with size-specific prey consumption rate 5 to 18 times the rate of mysids. Results show that the studied planktivores are capable of adjusting their feeding behaviour to decrease their conspicuousness in order to increase survival under predation risk. Further, results support the view that C. pengoi has adapted well to the Baltic ecosystem, sharing food niche with pelagic mysids and most probably having a strong influence on the whole pelagic food web.     

Effects of acute changes in temperature and salinity on the oxygen uptake of larvae of herring (Clupea harengus) and plaice (Pleuronectes platessa)

Routine oxygen uptake (QO₂) by yolk-sac and firstfeeding larvae of herring (Clupea harengus L.) and plaice (Pleuronectes platessa L.) was studied after acute change of temperature (8°, 13°, 18°C) and salinity (5, 12.7, 32, 40). In both species, QO₂ (l mg^-1 dry wt h^-1) of both larval stages increased with increasing temperature. Salinity effect on QO₂ varied: for yolk-sac larvae of both species a lower QO₂ was found at lower combined salinities (5 and 12.7); for feeding larvae a lower QO₂ was observed at 12.7 for both species, possibly due to the relatively smaller size of larvae used at this salinity. For both species, oxygen uptake increased as larvae grew and weight regression coefficients were between 0.74 and 1.33. At 32 S, no difference was found in oxygen consumption between species as a function of temperature.Based on a dissertation submitted in partial fulfillment of the requirements for the degree of Master of Science at the University of Stirling, Stirling, Scotland. The work was performed at the Dunstaffnage Marine Research Laboratory, Oban, Scotland     

A laboratory study of predation by Aurelia aurita on larval herring (Clupea harengus): Experimental observations compared with model predictions

Predation by the medusa Aurelia aurita L. on early first-feeding stage larvae of the herring clupea harengus L. was studied in the laboratory. The medusae were captured in Loch Etive, Scotland. Herring larvae were reared from the extificially fertilized eggs of spawning Clyde herring caught in March, 1982. Swimming speeds, volume searched”, capture efficiency and predation rates increased as medusa size increased. Predation rates on fish larvae increased with prey density, but appeared to approach a maximum at high prey densities; in 1 h experiments, a maximum rate of predation of 6.64 larvae h^-1 was estimated by fitting an Ivlev function. A model to predict predation rates was constructed from swimming speeds, sizes and densities of medusae and larvae, and capture efficiency. The rates of predation predicted from the model fell within the range of experimental data, but tended to underestimate rates and did not account for saturation of medusae. Swimming patterns of medusae changed after prey capture: (a) before capture, encounter rates were low and medusae were relatively less active; (b) after capture of 1 larva, encounter rates doubled, with the stimulated medusae exhibiting increased activity and an aftered “searching” path; and (c) after capture of many larvae, swimming speeds and encounter rates of medusae decreased.     

In-situ observations of Baltic herring (Clupea harengus membras) spawning behaviour in the Askö-Landsort area,northern Baltic proper

In a Baltic herring (Clupea harengus membras L.) spawning ground study, carried out in summer 1982 in the northern Baltic proper, spawning herring were encountered on two line transects on June 14. The spawning behaviour of minor groups as well as of a large school was studied in situ by four divers for approximately three hours. No behavioural differences or systematic coordinated behaviour between the sexes could be detected. Most of the spawning took place in a limited depth interval, 0.5 to 4 m, and Ceramium tenuicorne was the substrate most used.     

Effects of a transitory,low food supply in the early life of larval herring (Clupea harengus) on mortality,growth and digestive capacity

In order to assess possible effects of a transitory, low food supply on later development, three groups of Clyde herring larvae (Clupea harengus L.) were exposed in 1989 to different feeding regimes immediately after yolk resorption. Group 1 received a high daily ration of 80 copepods larvae^–1 for 31 d, Group 2 a low daily ration of 15 copepods larva^–1 for 10 d followed by a high ration (80 copepods larva^–1) for 21 d and Group 3 a low ration of 15 to 20 copepods larva^–1 for 31 d. After 31 d of feeding, digestive capacity, expressed as the sum of trypsin and trypsinogen, was markedly reduced in Group 2 compared to Group 1, while Group 3 had an even lower digestive capacity. After the switch from low to high ration Group 2 exhibited compensatory growth and caught up with Group 1 both in standard length and content of soluble protein. Group 3 had the lowest growth rates. Mortality was equal in Groups 1 and 2, while Group 3 showed an excess mortality of 40% of the start population. Although Group 2 larvae had caught up with Group 1 in growth at the end of the study, content of trypsin and trypsinogen in Group 2 was only half of that found in Group 1. Thus, comparing effects of a short period of food limitation on future growth, mortality and content of digestive enzymes, the study indicates content of trypsin and trypsinogen to be the most sensitive variable for detection of food limitation in the early stages of exogenous feeding.     

Quantification of trypsin with a radioimmunoassay in herring larvae (Clupea harengus) compared with a highly sensitive fluorescence technique to determine tryptic enzyme activity

Enzymatic activity and quantity of the protease trypsin were measured in individual herring larvae (Clupea harengus L.). The enzymatic activity assay was done using a fluorescence technique, and a radioimmunoassay was used for quantification of trypsin. The results are compared and the differences between the techniques discussed. Both methods gave similar results, as high or low values in trypsin quantity were reflected in high or low values of tryptic activity. Quantity and activity were linearly and positively correlated, but small differences between methods were found at the lowest detection limits. Both techniques reflect high variability between individual larvae.