Trypsin content in intestines of herring larvae,Clupea harengus,ingesting inert polystyrene spheres or live crustacea prey

In 1986, at the Danish Institute of Fisheries and Marine Research, Denmark, Clupea harengus L. larvae from three different herring stocks were offered either non-biodegradable polystyrene spheres, nauplii and copepodites of Acartia tonsa or Artemia ssp. nauplii. Ingestion of polystyrene spheres induced trypsin secretion to a higher level than in non-feeding fish. Larvae ingesting live food of the same width as the polystyrene spheres exhibited the highest trypsin content in the intestines. Mechanisms responsible for the regulation of pancreatic enzyme secretion are discussed.     

Prey selection by larval weakfish (Cynoscion regalis): the effects of prey size,speed, and abundance

We examined feeding by larval weakfish, Cynoscion regalis (Bloch and Schneider), in laboratory experiments conducted during the 1991 spawning season. under natural conditions weakfish larval development is ca. 3 wk, and we ran separate experiments with larvae of five different ages (5, 8, 11, 14, and 17 d post-hatching). We used two different size classes of rotifers (Brachionus plicatilis) and brine shrimp nauplii (Artemia sp.) as prey organisms. Contrary to results of previous research, weakfish larvae did not select prey based on size alone. When prey abundance was above 100 itemsl^-1 weakfish, larvae always chose large rotifers (length = 216 m) over small rotifers (length = 160 m). At 11 d post-hatching, larvae switched their diet from large rotifers to small brine shrimp nauplii (length = 449 m); however, when fed small rotifers and small brine shrimp nauplii the change in diet occurred at 14 d post-hatching. This pattern of selectivity was maintained in each larval age class. Early-stage larvae (5 and 8 d post-hatching) did not feed selectively when prey abundance was less than 100 itemsl^-1. Late-stage larvae (17 d post-hatching) fed selectively at abundances ranging from 10 to 10000 items^-1. Lwimming speeds of prey items, which ranged from 1 to 6 mms^-1, had no consistent effect on prey selection. These results suggest that weakfish larvae are able to feed selectively, that selectivity changes as larvae age, and that selectivity is also influenced by prey abundance.     

Feeding behaviour and prey size selection of gilthead seabream,Sparus aurata,larvae fed on inert and live food

Prey selection shortly after the onset of feeding by laboratory-reared gilthead seabream, Sparus aurata L., larvae was studied using larvae fed on two types of microcapsule (hard- and soft-walled) having diameters ranging from 25 to 300 m. Preferences between inert food and live prey (rotifers and Artemia sp. nauplii) were also studied. Seabream larvae were able to ingest inert food from first feeding. Larvae of all size classes ingested hard microcapsules with diameters in the range 25 to 250 m. However, larvae with a total length (TL) below 4 mm preferentially selected particles 25 to 50 m in diameter, larvae of TL 4 and 5 mm preferred particles 51 to 100 m in diameter, while larvae above 5 mm TL preferred particles 101 to 150 m in diameter. With soft microcapsules, larvae always preferred particles larger than in the previous case, and above 4.5 mm TL they preferentially selected particles 201 to 250 m in diameter. In addition, the gradual increase of preferred diameters with increasing TL was more pronounced when larvae were increasing TL was more pronounced when larvae were fed on soft particles. Mean values for prey width/mouth width ratios were approximately 0.24 and 0.30 when larvae were fed on hard-walled and soft-walled microcapsules, respectively, irrespective of the absolute value of larval length. When a mixed diet of live and inert food items was offered, live prey were always preferentially selected, even if the prey width/mouth width ratio was apparently not favourable. Only a physical constraint such as excessive prey width could counter this preference for living prey vs inert microcapsules. These results contribute to our knowledge in larval feeding behaviour, especially in the presence of inert food, and represent a fundamental step in developing prepared food for marine fish larvae.     

Effects of copper and cadmium on growth,swimming and predator avoidance in Eurytemora affinis (Copepoda)

In this study we demonstrate the sensitivity of swimming behavior and predator-escape responses of nauplii of the estuarine copepod Eurytemora affinis to sublethal doses of Cu and Cd. Behavior was generally altered at metal doses below those affecting growth rates or survival of the copepods. Swimming velocities of Cu-dosed nauplii were different from controls at all concentrations of Cu tested (10–50 g l^-1 total Cu) after 24- to 48-h exposure, whereas development rate of nauplii was significantly reduced only after 96 h at 25 g l^-1. The 96 h LC50 for Cu was approximately 30 g l^-1 Cu. Naupliar swimming velocity was also affected by Cd. Swimming speeds were reduced after 24 h at 130 g l^-1, and development was slowed after 48 h at 116 g Cd l^-1. The 96-h LC50 was >120 g l^-1. Little is known of the adaptive role of specific motile behaviors in the success of larval copepods. We investigated the relationship of swimming speed to predator — prey interactions of the nauplii using both real and simulated predators. Nauplii exposed to Cu for 24 h were observed to be generally hyperactive, a condition which could increase their encounter frequency with predators. Reduced numbers of escape responses of nauplii to a simulated predator, another indication of increased vulnerability to predation, were observed only after 48-h exposure to Cu. Nevertheless, feeding rates of non-dosed larval striped bass on dosed nauplii (24 h at 25 g Cu l^-1) were significantly higher than on control nauplii. Feeding rates of larval mysid shrimp, however, were not higher on similarly dosed nauplii; 24 h exposure of nauplii to >30 g Cu l^-1 did result in increased predation by mysids.Contribution No. 272 of the US EPA Environmental Research Laboratory, Narragansett, Rhode Island 02882, USA     

Selective particle ingestion by oyster larvae (Crassostrea virginica) feeding on natural seston and cultured algae

I investigated selective particle ingestion by oyster larvae (Crassostrea virginica) feeding on natural seston from Chesapeake Bay and laboratory-cultured algae of different sizes or chemical content. In 15 of 16 experiments with complex natural suspensions as food, small(<150 m) and large (>150 m) larvae selected most strongly for small (2 to 4 m) food particles, but in the presence of a large (>10 m)-cell dinoflagellate bloom, large larvae strongly selected much larger (22 to 30 m) food material (presumably dinoflagellates). When fed simplified mixtures of four cultured algal species (Synechococcus bacillaris, Isochrysis sp., Dunaliella tertiolecta, and Prorocentrum minimum) ranging in size from 1 to 11 m, small larvae preferred 1 m algae while large larvae preferred 11 m algae. In experiments with algal mixtures, and with suspensions of natural particles and added algae, large larvae preferred algal species harvested from exponential-phase cultures over other species from stationary-phase cultures. Larval ingestion rates of the cultured alga Thalassiosira pseudonana were about three times higher for cells with a low carbon:nitrogen ratio (7.2:1) than for high C:N ratio (16.2:1) cells when these cells were offered separately in suspensions of equal concentration. As a result, more algal cells, algal C, and algal N was ingested by larvae fed low C:N cells. However, larvae did not show a significant preference for either type of cell when they were offered in a 1:1 cell mixture. Feeding patterns of C. virginica larvae in natural food suspensions can vary with the composition of these complex suspensions, and ingestion seems dependent not only on the size, but on the growth rate and chemical quality of food particles.     

Food selection by laboratory-reared larvae of the scaled sardine Harengula pensacolae (Pisces,Clupeidae) and the bay anchovy Anchoa mitchilli (Pisces,Engraulidae)

Food selection by laboratory-reared larvae of scaled sardines Harengula pensacolae, and bay anchovies Anchoa mitchilli, was compared. Natural plankton was fed to the larvae during the 22 days following hatching. Food levels in the rearing tanks were maintained at an average of 1,600 to 1,800 potential food organisms per liter. Larvae of both species selected as food copepod nauplii, copepodites, and copepods; initial feeding was on organisms of 50 to 75 body width. Larvae of H. pensacolae averaged 4.2 mm in total length at hatching and those of A. mitchilli about 2.0 mm. H. pensacolae larvae grew about 1.0 mm per day and A. mitchilli 0.70 mm per day. The mean number of food organisms in each digestive tract was greater in H. pensacolae than in A. mitchilli, and the difference in number increased as the larvae grew. Average size of food organisms eaten increased for both species with growth, because of selection by the larvae; the average size of copepodites and copepods in digestive tracts increased at a faste rate in H. pensacolae than A. mitchilli. A. mitchilli longer than 8 mm did not eat copepod nauplii.Contribution No. 170, Bureau of Commercial Fisheries Tropical Atlantic Biological laboratory, Miami, Florida 33149, USA.     

Essential fatty acid (docosahexaenoic acid, DHA) availability affects growth of larval herring in the field

Larval fish growth and survival depends not only on prey quantity, but also on prey quality. To investigate effects of prey fatty acid concentration on larval herring growth, we collected different prey organisms and larval herring (Clupea harengus L.) in the Kiel Canal during the spring season of 2009. Along with biotic background data, we analysed fatty acids both in prey organisms and in the larvae and used biochemically derived growth rates of the larvae as the response variable. Larval herring reached their highest RNA/DNA derived growth rates only at high docosahexaenoic acid (DHA) concentration. When the ratio of copepodids to lesser quality cirriped nauplii was low, larval growth and larval DHA concentration were both significantly negatively affected. This was true even as prey abundance was increasing. This finding indicates that even in mixed, natural feeding conditions, growth variations are associated with DHA availability in larval fish.     

Diet of 0-group stages of capelin (<Emphasis Type="Italic">Mallotus villosus</Emphasis>), herring (<Emphasis Type="Italic">Clupea harengus</Emphasis>) and cod (<Emphasis Type="Italic">Gadus morhua</Emphasis>) during spring and summer in the Barents Sea

Recruitment of capelin in the Barents Sea fail when juvenile herring and cod are abundant and the potential for feeding competition of wild sympatric capelin and herring larvae and small cod juveniles were investigated. The frequency of gut evacuation after capture of capelin larvae were also studied in mesocosms. Small capelin larvae (<35 mm length) fed on small prey including phytoplankton, invertebrate eggs and nauplii, bivalves, other invertebrate larvae and small copepods. Calanus copepodites were only observed in large capelin larvae (>26 mm length). Calanus copepodites were the major food sources for contemporary herring larvae (25–35 mm length) and Calanus and euphausiids were the major prey for small juvenile herring (37–60 mm length) and cod (18–40 mm length). Capelin larvae reared in mesocosms evacuated the guts shortly after capture. Capelin larvae had a smaller mouth and fed on smaller prey than herring and cod of the same length. This implies that the small capelin larvae, in contrast to sympatric small herring and cod, are not tightly linked to the food chain involving Calanus and euphausiids. Thus, exploitative competition between capelin larvae and planktivorous fish that rely on Calanus and euphausiids in the Barents Sea may be relaxed.     

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.     

Formation of initial daily increments in sagittal otoliths of reared and wild Sardina pilchardus yolk-sac larvae

Daily deposition of growth increments in sagittal otoliths of reared and wild Sardina pilchardus (Walbaum, 1792) larvae from hatching to complete yolk-sac absorption is reported. Increments laid down prior to hatching were observed. A distinct growth increment, 1 m wide and located 5 to 7 m from the focus is laid down on the day of hatching. Subsequently, narrower increments (<1 m) are laid down daily throughout the yolk-sac stage. Measurements were made on the maximum radius of the sagittal otoliths of two groups of larvae: 83 larvae reared from fertilized eggs sampled from Ría de Muros (NW Spain), and 239 wild individuals sampled along the northern coasts of the Iberian peninsula. Counting of daily ring increments can provide a direct and valid estimation of larval age without the necessity for later correction.     

Acute toxicity of mercury and selenium to Crassostrea gigas embryos and Cancer magister larvae

The possible modification of mercury toxicity by selenium in embryos of the Pacific oyster Crassostrea gigas and the larvae of the crab Cancer magister was investigated. Mercury concentration eliciting abnormal development in 50% of the oyster embryos (EC₅₀) was 5.7 g l^-1 (48 h) and mortality in 50% of the crab larvae (LC₅₀) occurred with 6.6 g l^-1 (96 h). The 48 h EC₅₀ for selenium was greater than 10,000 g l^-1 for oyster embryos and the 96 h LC₅₀ for crab zoeae was 1040 g l^-1. The response from each species, when exposed to both toxicants, revealed, that a high level of selenium (5,000 g l^-1) increased mercury toxicity. Moderate selenium concentrations (10 to 1,000 g l^-1) tended to decrease mercury toxicity, although no statistical verification could be made. The order of administration of toxicants had no effect on the response of Crassostrea gigas embryos. Early developmental stages (8 h) of C. gigas embryos were most sensitive to dissolved Hg; toxicant administration 24 h after fertilization resulted in no apparent abnormalities in development.     

Osmoacclimation in Enteromorpha intestinalis: long-term effects of osmotic stress on organic solute accumulation

The fate of the protease trypsin in intestines of individual herring larvae Clupea harengus L. was studied following digestion of the copepod Acartia tonsa. Trypsin was retained in the intestine during two consecutive pulses of feeding and defaecation of copepods. Quantification of herring trypsin in digested, defaecated copepods showed that ca. 1% of larval intestinal enzyme was defaecated along with 1 to 3 copepods. Following ingestion of a single meal, the level of intestinal trypsin post-ingestion declined to pre-ingestion levels within 1 to 2 d of starvation. All enzyme data thus indicated that trypsin, released in response to ingestion of a meal, was retained. In addition, analysis of fed subgroups of starved larvae clearly indicated that release of trypsin from the pancreas stopped after 6 to 8 d of starvation. As the fish still contained substantial amounts of trypsinogen, the underlying cause might be defective release mechanisms. Daily secretion of trypsin and processes responsible for enzyme retention in the gut are discussed. Assimilation efficiency in herring larvae was estimated for copepodite prey. Average carbon assimilation was 90%.     

Daily ration and specific daily ration of the chaetognath Sagitta enflata

Digestion times, obtained directly from laboratory feeding, were used in conjunction with a day/night series of field samples to estimate the daily ration and specific daily ration of Sagitta enflata Grassi, the most abundant chaetognath in the Gulf Stream near Miami, Florida (USA), during winter and early spring. Feeding was independent of time of day and increased with increasing chaetognath size from 1.7 prey day^-1 for the 11.5 mm size class to 2.9 prey day^-1 for the 21.5 mm class. The diet of s. enflata consisted of 94.8% copepods having a mean weight of 21.8 g, and 5.2% prey chaetognaths estimated to be two-thirds the length of the predators. The daily ration of S. enflata from 12.5 to 20.5 mm in length increased from 48.3 to 143.3 g of prey dry weight and from 18.5 to 48.1 g of prey carbon. The specific daily ration declined from 0.12 to 0.08 dry weight basis, and from 0.26 to 0.14, carbon basis, over the same size range. Prey chaetognaths contributed as much as 51% of the ration on a carbon basis.     

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.     

Larval development in the Antarctic nemertean Parborlasia corrugatus (Heteronemertea: Lineidae)

Embryonic and larval development were followed from fertilisation to settlement in the Antarctic heteronemertean Parborlasia corrugatus (McIntosh, 1876). The first cleavage occurred 10 to 15 h after fertilisation, and the second at 17 h. Larvae hatched at the gastrula stage, between 170 and 200 h post-fertilisation, and were 150 m in diameter. Early larval stages aggregated in dense groups near the surface of incubation vessels and were positively phototactic. Early pilidium larvae were recognisable 435 h post-fertilisation. They were 155×152 m in size, and possessed a complete apical tuft of cilia and a full marginal band of locomotory cilia. At this stage, the gust was visible through the body wall, and the mouth was open and was 40 m in diameter. Late pilidia, 222×193 m in size, were helmet-shaped. They had an apical tuft over 100 m long, and possessed a lobed marginal band of locomotory cilia. Pilidia were observed aggregating close to the bottom of incubation vessels 1200 to 1350 h (50 to 56 d) after fertilisation, and this was interpreted as settlement behaviour. At this stage, the apical tuft had been lost and they were highly contractile, being capable of compressing their bodies. However, neither developing juveniles within the larval envelope nor hatched juveniles were observed. Pilidia consumed the microalgae Tetraselmis suecica, Thalassiosira pseudonana and Isochrysis galbana. They also fed on particulate organic material < 1 m in size, as shown by the presence of material in the guts of larvae offered filtered extracts of algal cultures. There was some indication that larvae could use dissolved organic material, since pilidia held in seawater with organic material removed did not survive as long as those in filtered seawater or in filtered water with added amino acids. However, the only larvae to exhibit settlement behaviour in the feeding experiments were those offered Tetraselmis succica and Thalassiosira pseudonana, and these required a longer development time to reach this stage than pilidia in the standard cultures, where a mixed algal diet was offered.     

Growth and survival of Mytilus edulis larvae exposed to low levels of dibutyltin and tributyltin

Two studies were conducted to observe effects of dibutyltin (DBT) and tributyltin (TBT) on larvae of Mytilus edulis for an exposure period of 25 d. Endpoints for evaluation were shell growth and mortality measured at 33 d. Larvae were cultured in a new laboratory assay chamber in a recirculating static test. The control, 2, 20, and 200 g/l DBT-treated populations had mean shell lengths of 527, 523, 417, and 180 m, respectively. Survival was 1% for the 200 g/l DBT-treated population, but ranged from 73 to 83% for controls, 2, and 20 g/l treatments. The no-observed-effect concentration (NOEC) was 2 g/l for DBT, while the lowest-observed-effect concentration (LOEC) was 20 g/l. The chronic toxicity value was 6.3 g/l. In the TBT bioassay, mean shell lengths for the control, 0.006, 0.050, and 0.130 g/l-treated populations were 565, 437, 385, and 292 m, respectively. Control survival was 74%, whereas TBT-treated populations survival ranged from 52 to 58%. The NOEC for TBT was 0.006 g/l TBT and the LOEC was 0.050. A chronic toxicity value of 0.017 g/l was calculated. The results of this study indicated that the toxicity of DBT was less than that of TBT. It was concluded that shell length was inversely related to exposure level in both DBT and TBT bioassays. In this study, we have observed TBT effects at lower exposure levels in the laboratory than previously reported, and also report the first data for DBT effects on mussel larvae.     

Effect of cadmium on survival and osmoregulation of various developmental stages of the shrimp Penaeus japonicus (Crustacea: Decapoda)

The object of this study was to evaluate the acute toxicity of cadmium in different post-embryonic stages of the penaeid shrimp Penaeus japonicus (Bate, 1888) and to determine the effect of sublethal cadmium on the osmoregulatory capacity used as an indicator of physiological condition. Tolerance to cadmium increases with the developmental stage. The least tolerant stages are the nauplii (48 h LC₅₀: 124 g Cd l^-1) and the zoeae (96 h LC₅₀: 10 to 30 g Cd l^-1). The most tolerant stages are the postlarvae (96 h LC₅₀: 200 to 3500 Cd l^-1) and juveniles (96 h LC₅₀: 5500 g Cd l^-1). In juvenile shrimp, 2000 g Cd l^-1 significantly reduce hypo- and hyper-OC. The effect of cadmium on hypo- and hyper-osmoregulatory capacity illustrates a dose- and time-dependent response. Surviving shrimp recover their hypo-osmoregulatory capacity after 6 d of readaptation in cadmium-free seawater.     

Mass culture of the rotifer Brachionus plicatilis and its evaluation as a food for larval anchovies

Growth rates of anchovy larvae, Engraulis mordax, reared for 19 days under constant environmental conditions on a diet of laboratory-cultured organisms, exceeded the growth rates of anchovies fed on a diet of wild plankton. The rotifer Brachionus plicatilis was found to be a nutritous food source when fed to the larvae in concentrations of 10 to 20/ml and in combination with the dinoflagellate Gymnodinium splendens (100/ml). Optimum conditions were determined for mass culture of the rotifer. A high food concentration was the most important parameter needed to assure a high yield of rotifers. Large volumes (464 I) of the unicellular flagellate Dunaliella sp. were cultured for feeding the rotifers. The rotifer culture technique described produces approximately 2.5×10⁶ organisms/day, providing a reliable food source for rearing studies. The lengths of B. plicatilis (without eggs) ranged between 99 and 281 , most rotifers being larger than 164 and less than 231 . Individuals weighed 0.16 g and contained 8×10^-4 cal.     

Sublethal effects of zinc and municipal effluents on larvae of the red abalone Haliotis rufescens

Experiments were conducted to develop a sensitive sublethal toxicity test protocol to determine the toxicity of municipal wastewater effluents to larvae of the red abalone Haliotis rufescens. In multiple tests, fertilized abalone embryos were exposed for 48 h to dilutions of a reference toxicant, zinc sulfate, and to dilutions of primary-and secondary-treated effluents. The resulting veliger larvae were examined microscopically for larval shell abnormalities. In a longer flowthrough experiment, abalone were exposed for the entire larval phase, from the two-cell stage through metamorphosis, to compare zinc effects on metamorphosis with zinc effects on short-term larval shell development. Dissolved oxygen, pH, salinity and temperature were measured daily in test solutions, and zinc concentrations were verified by chemical analysis. No observed effect concentrations (NOECs) for zinc were 39±2.1 g l^-1 in three 48 h exposures, and 19 g l^-1 for the 9 d exposure through metamorphosis. Median effect concentrations (EC₅₀s) were 68±6.9 g l^-1 in 48 h tests and 50 g l^-1 in the 9 d test. Abalone larvae were affected at lower concentrations of primary than of secondary effluent.     

Morphological and histological changes during the growth and starvation of herring and plaice larvae

Reared herring (Clupea harengus L.) and plaice (Pleuronectes platessa L.) were examined for morphological and histological changes during growth and starvation. The growth rate of herring larvae of 0.22 mm/day was less than that reported for wild stock, but this difference was attributed to survival of runts in laboratory. Larval plaice had a growth rate of 0.16 mm/day. The relative condition factor (antilogarithm of intercept of length-weight line) was used to assess condition throughout the larval stages. Starvation resulted in a progressive collapse of the larval body, especially of the ventral body surface around the pectoral girdle of both species (assessed by the pectoral angle) and of the spacing between the organs of the head in herring. There was a breakdown of the herring gut with decreases in epithelial cell height and catabolism of the connective tissue coat and a marked reduction in the transverse sectional area of the plaice liver. The changes in the pectoral angle in both herring and plaice and the eye height to head height ratio in herring should be useful to fishery biologists for assessing nutritional condition, even on board ship.