Changes in nucleic acids over the molt cycle in relation to food availability and temperature in Homarus americanus postlarvae

Postlarval lobsters Homarus americanus Milne Edwards hatched from three females collected in 1989 from Block Island Sound, Rhode Island were reared individually in the laboratory under nine treatment combinations of temperature (15, 18 and 21°C) and feeding (starved, low ration and full ration). Total RNA, DNA (mgind.^-1), RNA:DNA ratios and molt stage were determined for individuals sampled at daily intervals. Postlarval lobsters had high resistance to starvation. A majority of the lobsters survive 12 d of food deprivation, with some surviving up to 24–29 d. During starvation, cell biomass (estimated from protein:DNA) decreased to a minumum size, whereas cell number (based on total DNA) was generally conserved. The molt cycle was arrested at molt stage C in the starved postalrvae. Instar duration was inversely related to temperature. However, the duration of the postlarval instar did not differ between the low and full ration treatments. Uncoupling of cell growth and the molt cycle was evident in the full and low ration treatments. In the full ration treatments, the postmolt through early premolt was characterized by a rapid increase in total RNA and DNA. Maximum cell biomass was attained by molt stage C or D₀. In the low ration treatments, total RNA and DNA were less than those in the full ration treatments and the maximum cell biomass was attained only at molt stage D_2-3. Notably, different feeding regimes resulted in different patterns in RNA:DNA ratios over the molt cycle during the postlarval instar. RNA:DNA ratio was a sensitive indicator of recent (2 to 3 d) food deprivation. However, this ratio was positively related to the level of feeding only at molt stages C to D₁ and was inversely related to temperature regime. These results indicate that the use of RNA:DNA ratios to estimate the relative nutritional state of postlarval tobsters must be qualified with respect to the period of the molt cycle and the temperature regime.     

Biochemical responses during starvation and subsequent recovery in postlarval Pacific white shrimp,Penaeus vannamei

Postlarval shrimp, Penaeus vannamei Boone, 1931, were held individually in cages and exposed to two feeding regimes. One group was starved for 12 d and then fed during the following 12 d. A second group was fed throughout the 24 d study. Four individuals were sampled from each of the two groups on Days 0, 1, 2, 4, 8, 12, 13, 14, 16, 20, and 24. Molting and growth among the starved-fed postlarvae stopped after 2 d starvation, while fed postlarvae increased significantly in size throughout the 24 d study. Among the starved-fed postlarvae, water content increased rapidly in response to starvation. DNA and sterol concentrations increased significantly during starvation due to selective catabolism of cellular components. After 12 d, RNA concentration was not significantly different between the fed and starved-fed postlarvae, but became significantly higher in the starved-fed postlarvae 48 h after feeding resumed. Triacylglycerol reserves were severely depleted during the first day of starvation, while protein concentrations began to decrease after the second day of starvation. RNA, protein, and the polyamines spermidine and spermine, when expressed as a ratio to DNA, decreased in response to starvation. Concentrations of all measured parameters in the starved-fed postlarvae returned to levels similar to those in the fed group 8 to 12 d after feeding resumed. Results of this study suggest that triacylglycerol provides energy during short periods of starvation, while protein is utilized during prolonged starvation. The ratios of RNA:DNA, protein:DNA, spermidine:DNA, spermine:DNA, two unidentified amine compounds, and percent water content are all useful indicators of prolonged nutritional stress in postlarval P. vannamei.     

Biochemical changes during larval development of the xanthid crab Rhithro panopeus harrisii. II. Nucleic acids

Deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) contents were measured daily during the zoeal development of the crab Rhithropanopeus harrisii (Gould). DNA concentration (per unit protein) decreased as larvae increased in size. Total DNA content per larva showed an abrupt increase at the second molt and increased steadily during the third and fourth zoeal stages. Ratios of fresh weight to DNA were highest during the first two zoeal stages, dropped sharply at the second molt, then increased during the third and fourth zoeal stages. RNA:DNA ratios showed cyclical activity apparently related to the molt cycle. Assuming that fresh weight: DNA ratio reflects cell size and that DNA content reflects cell number, growth during the first two stages is due primarily to increase in cell size and during the last two stages to a combination of increase in cell size and number. Assuming further that RNA:DNA ratios reflect protein-synthesis activity, such activity in these larvae follows a rhythm related to the molt cycle.     

Size regulation in larvae of the crustacean Balanus eburneus (Cirripedia: Thoracica)

We tested the hypothesis that larval size in the acorn barnacle Balanus eburneus Gould (Cirripedia Thoracica) varies in relation to food availability. In March–November 1980, and March–July 1981, larvae were obtained from adult Balanus eburneus collected in the Newport River, North Carolina, USA. Carapace length and width of larvae reared at three different food concentrations were measured. Mean naupliar instar size was independent of food concentration. Mean size of the cypris instar increased with increasing food level. Greater cypris size could be attributed to increased food reserves in the preceding naupliar stage, and was coinciden with inmarked increase in metamorphic success. Variation in instar size remained constant or declined during naupliar development, but increased sharply at the molt to the cyprid. Naupliar size regulation involved: (1) conservation of a molt increment specific for each naupliar-naupliar molt, (2) an inverse relationship between premolt size and the molt increment during the first five naupliar instars, and (3) an increase in the precision of the molt increment at the molt to the sixth naupliar instar. Experimental evidence implies that size regulation in Balanus eburneus limits variation about a fixed final naupliar size (e.g. volume). Measurement of naupliar size, accumulated energy reserves, survival and development time, and cypris metamorphic success indicated that naupliar cuticular growth is the most conservative feature of larval development. The data suggest that maximum naupliar size is limited by escalating metabolic costs during development, while minimum naupliar size is limited by size-related feeding effectiveness.     

Measuring RNA:DNA ratios in individual <Emphasis Type="Italic">Acartia tonsa</Emphasis> (Copepoda)

Acartia tonsa Dana is a dominant copepod in coastal waters and is therefore an important link in the food web between microplankton and higher trophic levels. RNA:DNA ratios have been used to describe growth and nutritional condition of field-collected copepods and to show strong correlation between RNA:DNA ratios and group egg production (EP). A method was developed using a sensitive, nucleic acid-specific fluorescent dye, and automated microplate fluorometer to measure DNA, RNA, and the RNA:DNA ratio of individual A. tonsa. DNA, RNA, and RNA:DNA ratios and EP were all significantly higher in copepods fed Thalassiosira sp. compared to starved copepods. There was a general trend toward an increase in RNA:DNA ratios with increase in EP, but due to the high degree of variation in both RNA:DNA ratios and EP among individual copepods there was no significant correlation between RNA:DNA ratios and EP. Significant differences in RNA:DNA ratios between fed (7.2) and starved (3.3) copepods were found after 2 days. This assay may be applied to other species of copepods sampled in the field to provide an index of the health of planktonic food webs.     

Nucleic acids and growth of larval and early juvenile spider crab,Hyas araneus

The spider crabHyas araneus (L.) was collected from the North Sea in winter 1986–1987 and reared in the laboratory from hatching of the Zoea I (ZI) through the first juvenile instar (CI). Within a given moult cycle, individuals of the same age were sampled in intervals of 2 (ZI, ZII, CI) or 3 d (megalopa) for analysis of dry weight (W), carbon (C), nitrogen (N), hydrogen (H), protein, DNA, and RNA. Lipid was calculated from C. Biomass, growth rate and nucleic acid contents showed high variability during each moult cycle and between instars. Instantaneous growth rates of C were high in postmoult and intermoult, and low in the premoult period of each moult cycle. A shift was observed from high rates of lipid accumulation in the postmoult and intermoult stages to proportionally increasing protein accumulation during late premoult (ZI), or throughout a major part of the remaining moult cycle (in all other instars). DNA was accumulated throughout the ZI and ZII instars, but decreased in late premoult megalopa. It increased again from late intermoult through intermediate premoult in juveniles. RNA increased continuously during ZI and ZII, and decreased in the megalopa, almost to levels that had been found immediately after hatching. In juveniles, variation in RNA followed closely those in DNA. Cell multiplication (expressed by DNA increase) dominated over increase in cell size (defined by the C/DNA ratio) during the zoeal instars and in postmoult through early intermoult in the megalopa and CI. When specific (C-related) RNA values and RNA/DNA ratios were compared with instantaneous growth rates in C and N, no general correspondence was detected. The only significant relationship between specific RNA values and instantaneous C or N growth rates was found in the megalopa. The same held for the relationship between the RNA/DNA ratio and growth. Here, in addition to the megalopa, a correspondence with C growth was also found in the CI instar. Our results suggest that variation in nucleic acids may provide useful insights into mechanisms of growth on the cellular level (cell multiplication vs cell enlargement). However, lack of general correlation with variation in growth rates ofH. araneus larvae shows that the use of nucleic acids as a measure of growth is probably based upon too simplistic assumptions; it may not yield reliable predictions, when growth is associated with developmental events.     

Feeding ecology of the three juvenile phases of the spiny lobster<Emphasis Type="Italic"> Panulirus argus</Emphasis> in a tropical reef lagoon

The three juvenile phases of the spiny lobster Panulirus argus (algal phase: 5-15 mm carapace length, CL; postalgal phase: 15-45 mm CL, and subadults: 45-80 mm CL) occur in the reef lagoon at Puerto Morelos, Mexico. The algal phase abounds in this lagoon, which is covered by extensive seagrass-algal meadows, but the density of postalgal and subadult juveniles is low, owing to the scarcity of crevice-type shelters suitable for these phases. The feeding ecology of the three juvenile phases was investigated to examine whether spatial or temporal differences in food intake, diet composition, or nutritional condition occurred among phases and could partially account for the low abundance of the larger juveniles. Juveniles were collected by divers at night, from January to November 1995, throughout the mid-lagoon and back-reef zones. Percent stomach fullness, relative weight of the digestive gland (RWDG, an index of nutritional condition), percent frequency of occurrence and percent volume of food categories in the diet were compared between sexes, juvenile phases, molt stages (postmolt, intermolt, premolt), seasons, and sampling zones (mid-lagoon and back-reef zones). Significant differences in stomach fullness occurred only among molt stages, mainly because postmolt individuals had emptier stomachs. The main food categories in all juvenile phases were crustaceans (mostly hermit crabs and brachyurans) and gastropods, but the food spectrum was wide, including many other animal taxa as well as plant matter. In June 1995, the epibenthic macrofauna was sampled in five sites in the lagoon that differed in their amount of vegetation. The most abundant taxa in all sites were decapods and gastropods, but density and diversity measures showed that the distribution of these potential prey taxa for juvenile P. argus was rather patchy. Diet overlap in juvenile lobsters was high between sexes, juvenile phases, sampling zones, seasons, and molting stages, indicating that all juveniles fed on the same general food categories throughout time. The only factor that affected the RWDG was the juvenile phase. RWDG was significantly lower in subadults than in algal and postalgal phases, suggesting a poorer nutritional condition in the largest juveniles. This may be related to the scarcity of suitable shelters for large juveniles throughout the lagoon, which may preclude subadults from exploiting food resources in areas of the lagoon where shelter is limited.     

Study by evolution of nucleic acid content of prepuberal growth in the shrimp Crangon vulgaris

Growth of the shrimp Crangon vulgaris was studied by following the evolution of its nucleic acid concentration and total content. Deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) analysis were effected on homogenates of whole shrimps. Cell multiplication was estimated from evolution of total DNA content, and cell size increment from evolution of fresh weight: DNA ratio. DNA puric bases ratio and RNA monoribonucleotides percentages were constant throughout the investigation period. A decrease in DNA concentration was observed from 50 to 20 M/g dry defatted weight (DDP), when fresh weight increases from 40 to 570 mg. This decrease was more marked in the first half of the period studied (prepuberal phase). The DNA content in the whole shrimp increases with fresh weight without slowing down at sexual maturity. However, at the beginning of the prepuberal phase the cell-multiplication rate seems to be graduated rather than continuous; this is still to be explained. Cell size, calculated from fresh weight: DNA ratio, increases until the shrimp weighs 300 mg; beyond this weight, cell increment proceeds much more slowly. The weight gain of shrimps throughout the growth period studied is mainly attributable to hyperplasia — which occurs constantly — while hypertrophy insures only 21% weight increment in the largest shrimps. As protein content, RNA content is a linear function of shrimp weight. RNA:DNA ratio evolution is similar to that of cell size and protein content. We suggest that cells have attained a physiological balance when shrimps reach a weight of 300 mg.     

The use of RNA:DNA ratios to predict growth limitation of juvenile summer flounder (Paralichthys dentatus) from Delaware and North Carolina estuaries

Nutritional indices were used to develop biochemical correlates of feeding and growth rates for juvenile summer flounder, Paralichthys dentatus (Linnaeus), from North Carolina (NC) and Delaware (DE). Six parameters (Fulton's condition K=10⁴xweight/(length³), wet weight/dry weight, [protein], [RNA], [DNA], and RNA:DNA) were related to feeding and growth rates of fish from previously reported 10 to 14-d experiments at temperatures ranging from 2 to 20 °C with varying feeding levels (0 to 100% and libitum). RNA:DNA ratios were the best predictors of growth rates, but inclusion of a temperature term improved the relationship between RNA:DNA ratios and growth rate for Delaware fish. Feeding rates were poorly correlated with all parameters. RNA:DNA ratios of fish in the laboratory changed significantly within 1 d of starvation and refeeding at 16 °C. RNA:DNA of juvenile summer flounder collected from one site in Indian River Bay, DE and two sites in the Newport River Estuary, NC, between January and June 1992 were used to estimate in situ growth rates following settlement. Predicted growth rates in both estuaries were close to maximum (suggesting ad libitum feeding) until early May. Growth rates of juveniles from Delaware were <0% d^-1 from December through early March, and were higher (0.6 to 3% d^-1) from April through early June. However, growth rates of DE juveniles during May were <50% of maxinum. North Carolina juveniles had growth rates of 2 to 5% d^-1 from February through early April. Juveniles from one of the Newport River sites (a marsh habitat) were also severely growth limited (<20% of maximum) after April. Prolonged periods of sub-optimal growth may be important to survival and recruitment of juvenile summer flounder in northern mid-Atlantic estuaries. A model is presented which illustrates the potential impact that small changes in temperature and growth limitation can have on recruitment success in both delaware and North Carolina estuaries.     

RNA:DNA ratio during the critical period and early larval growth of the red drum Sciaenops ocellatus

Eggs from two separate spawning stocks of the red drum Sciaenops ocellatus (Linnaeus) were hatched, and the larvae were reared in the laboratory for 2 wk under closely controlled conditions. Total RNA, DNA, and soluble protein were measured in each population daily in triplicate pooled samples of larvae from each of three tanks. Growth rate in mm d^-1 was determined for each population at 2 d intervals. Growth rate explained 72 and 95% of the variation in the RNA:DNA ratios of the two populations individually, and 86% of the variation in the RNA:DNA ratio when data from the two populations were combined. The RNA:DNA ratio appeared to be most effective as an indicator of growth in rapidly growing larvae, and to lose some resolution when growth was intermittent. The rates of deposition of RNA, DNA, and protein into tissue were all highly correlated with growth rate and with each other. Mean population RNA:DNA ratios of red drum yolk-sac larvae decreased from Day 1 post-hatch until larvae initiated successful feeding behavior, and then increased steadily throghout the remainder of the experimental period. This pattern of change in the RNA:DNA ratios during the yolk-sac stage appears to be an intrinsic developmental pattern of red drum ontogeny. The lowest values for the RNA:DNA ratio were observed just prior to the initiation of feeding or during the critical period, indicating that red drum larvae experience a decrease in capacity for protein synthesis as they initiate feeding. Intrinsic variation in the RNA:DNA ratio during development suggests that caution be used when comparing the RNA:DNA ratios of yolk-sac larvae to a critical ratio calculated from Buckley's general model.     

RNA/DNA ratio and expression of 18S ribosomal RNA, actin and myosin heavy chain messenger RNAs in starved and fed larval Atlantic cod (Gadus morhua)

Levels of total RNA, total DNA, 18S ribosomal RNA (rRNA), poly(A) messenger RNA (mRNA), and two mRNAs coding for abundant myofibrillar proteins were estimated in laboratory-reared Atlantic cod larvae (Gadusmorhua Linnaeus) under conditions of feeding and starvation. DNA probes specific for cod 18S rRNA, β-actin mRNA and myosin heavy chain mRNA were developed. In two experiments on newly hatched larvae in fed and starved treatments, changes in 18S rRNA and mRNA were similar to changes in total RNA during the first weeks after hatching. RNA levels in fed and starved larvae in both experiments were stable, or increased, over the first 3 d after hatching, and then decreased to minima at 9 d. RNA levels increased after 9 d, with the degree and timing of the increase varying among the individual classes of RNA. Complete mortality of starved larvae in both experiments was observed shortly after 11 d, corresponding to exhaustion of endogenous yolk reserves. Total RNA content, RNA/DNA ratio, 18S rRNA levels, total mRNA pool, and actin and myosin heavy chain mRNA levels showed significant differences in fed and starved first-feeding larvae after yolk exhaustion. In another experiment with 3- to 4-week-old cod larvae, 18S rRNA levels were significantly lower in starved versus fed larvae after 3 d. Total RNA responded to feeding and starvation within a similar time as 18S rRNA and the mRNAs examined. Analysis of bulk nucleic acids using fluorometric dyes was simpler and faster than analysis of individual RNAs using hybridization probes, and provides valuable information on recent growth and condition of individual larvae. However, analysis of specific RNAs can provide information on expression of the corresponding genes and reveal the changes underlying trends seen in bulk RNA. Received: 9 February 1996 / Accepted: 7 June 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.     

Respiratory regulation and the role of the blood in the burrowing shrimp Callianassa californiensis (Decapoda: Thalassinidae)

Questions relating to oxygen extraction and hemolymph oxygen transport have been investigated in the burrowing shrimp Callianassa californiensis Dana. The relationship of oxygen consumption to oxygen tension was determined in intermolt and postmolt shrimp. Soft postmolt individuals had higher rates of oxygen consumption and a somewhat lower critical oxygen tension than intermolt specimens. Both oxygen-carrying capacity and hemocyanin content are somewhat lower in postmolt compared to intermolt shrimp. The measured gradient between postbranchial (pericardial sinus) and prebranchial (abdominal sinus) hemolymph oxygen content is small and highly variable, and is not affected by exposure to anoxic stress. Hemolymph pH and buffering capacity are high. The oxygen consumption-oxygen tension relationships and respiratory properties of the hemolymph are discussed in relation to the hypoxic environment associated with the habitat of these crustaceans.Supported in part by Institutional Sea Grant 04-3-158-4 and Public Health Service Grant HL 12326.Communicated by J.S. Pearse, Santa Cruz     

Effect of salinity on hemolymph calcium concentration during the molt cycle of the prawn Penaeus monodon

Prawns (Penaeus monodon) were obtained from ponds in Iloilo, Philippines, in 1984 and 1985 and maintained in salinities from 8 to 44. Total hemolymph calcium was largely affected by molt stage and less so by salinity. A sharp, transient increase in hemolymph calcium occurred 3 to 6 h postmolt, followed by an equally rapid decrease from 6 h postmolt to intermolt. This biphasis response was limited to prawns in 8, 20 and 32S; in 44S, hemolymph calcium remained the same throughout the sampling period. Peak concentrations of total calcium were greater in low (8 and 20S) than in high salinities. Salinity had no effect on the duration of molt cycle nor on time of occurrence of molt. Almost half of molting incidents occurred between 18.01 and 0.00 hrs, and one-third between 0.01 and 06.00 hrs.     

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.     

Tests of the mate-guarding hypothesis for social monogamy: does population density, sex ratio, or female synchrony affect behavior of male snapping shrimp (Alpheus angulatus)?

There are at least two mechanisms by which social monogamy in the absence of biparental care may evolve: as a form of territorial cooperation, in which one or both sexes benefits by sharing a territory with a partner, and as a form of extended mate guarding, in which males guard females through entire, and perhaps multiple, reproductive cycles. I examined the effects of population variables (density, sex ratio, female synchrony) on male pairing behavior in the snapping shrimp, Alpheus angulatus, to test the hypothesis that social monogamy in this genus has evolved as a result of selection on males for long-term mate guarding of females. There was no evidence that pairing behavior changes with differences in population density; in a natural population, there was a 1:1 relationship between the number in pairs and local population density. In a laboratory experiment, males altered their pairing behavior in response to manipulated differences in sex ratio. Males in female-biased sex ratios were significantly more likely to abandon recently mated females than males in equivalent sex ratios, though there was no significant difference in the duration of pairing or the number of times males switched females. Observations of shrimp maintained for an extended period in the laboratory revealed no evidence that females molt and become sexually receptive synchronously, which would reduce the likelihood that a searching male would encounter additional receptive females. These data suggest that sex ratio may have contributed to the evolution of social monogamy in snapping shrimp, but provide no evidence that population density or female synchronous receptivity have contributed to the evolution of social monogamy.     

Relationship between growth and biochemical indices in laboratory-reared juvenile Japanese flounder (Paralichthys olivaceus), and its application to wild fish

In order to estimate growth rates based on biochemical indices of the liver of wild Japanese flounder (Paralichthys olivaceus), juveniles were reared at six ration levels (0, 0.5, 2, 4, 6 and 8% body weight day⁻¹) in the laboratory for 14 days, and the relationship between their growth rates and biochemical indices (RNA/DNA, protein/DNA, triglyceride/DNA, phospholipid/DNA and cathepsin D activities) were determined. Positive and approximately linear relationships were seen between growth rates and the indices of RNA/DNA, protein/DNA and phospholipid/DNA. The triglyceride/DNA ratio decreased with increasing growth rates up to approximately 1% body weight day⁻¹, then increased linearly with increasing growth rates. There was no significant correlation between growth rates and cathepsin D activity, and the highest values were obtained in the starved fish. Compared with laboratory-reared specimens, wild specimens of similar sizes were found to have significantly larger livers. The RNA/DNA, protein/DNA and phospholipid/DNA ratios of wild specimens fell in a broad range between ration groups of reared juveniles. The protein/DNA ratios of wild specimens were low and outside the range of the reared juveniles at six ration levels. In contrast, the levels of cathepsin D activity of wild fish were highest compared to the reared fish. Estimated growth rates of wild fish from the RNA/DNA, protein/DNA and phospholipid/DNA regressions obtained from the rearing experiment were 1.66, −1.74 and 0.10% day⁻¹, respectively. Based on our results, the RNA/DNA index may be regarded as the most valid and reliable growth estimator. It is noted that the larger liver size, the lower liver protein/DNA ratio and the unexpectedly high level of cathepsin D activities of wild specimens found in this study may reflect the different metabolic conditions of fish reared in the laboratory compared to those collected in the field. Received: 29 February 2000 / Accepted: 26 August 2000     

Seasonal changes in nucleic acids,amino acids and protein content in juvenile Norway lobster (<Emphasis Type="Italic">Nephrops norvegicus</Emphasis>)

The objective of this study was to describe the seasonal variations in nucleic acid contents and amino acid profiles in the muscle of juvenile Nephrops norvegicus. RNA and protein contents, and RNA:protein and RNA:DNA ratios varied significantly between seasons, being highest in spring and lowest in autumn/winter ( P<0.05). Though DNA content increased significantly from autumn to summer ( P<0.05), protein:DNA ratio did not show significant seasonal variations ( P=0.05). In respect to protein-bound amino acid content (BAA), a significant increase was observed from winter to summer ( P<0.05). Both essential (EAA) and non-essential amino acids (NEAA) increased significantly (27.6% and 27.8%, respectively; P<0.05), mainly due to the considerable increase in arginine and proline (59.1% and 225.2%, respectively; P<0.05). A significant decrease was observed in the free amino acid content (FAA) from winter to summer ( P<0.05); and a higher percentage decrease occurred in free non-essential (FNEAA; 27.9%) in comparison to free essential amino acids (FEAA; 21.8%). The significant increase in RNA and BAA contents from winter to spring may be related to protein synthesis. On the other hand, the lowest values obtained in winter may be due to a reduction in feeding activity; in this period the muscle protein must be progressively hydrolysed, which is evident with the higher FAA content. The liberated amino acids enter the FAA pool and become available for energy production.     

Condition of larval and juvenile red drum (Sciaenops ocellatus ) from estuarine nursery habitats

RNA:DNA ratios of larval and juvenile red drum (Sciaenops ocellatus) collected from nursery habitats in the Aransas Estuary, Texas, in 1994 were quantified using a highly sensitive ethidium-bromide fluorometric technique. RNA:DNA ratios of wild red drum were evaluated by comparing individual values to a linear regression model derived for starved laboratory-reared red drum. Wild red drum were in relatively good condition with <5% of the RNA:DNA ratios within or below the 95% prediction interval of 4 to 5 d starved red drum. A multiple-regression model explained 54% of the variability in the RNA:DNA ratio of wild red drum, and identified length and water temperature (midday) as significant factors. RNA:DNA ratios increased with fish length [≃1.1 mm⁻¹, over the size range investigated (5␣to 20 mm)]. The effect of temperature on the RNA: DNA ratio was assessed on different sampling trips, and ratios increased with increasing temperature. Abundance of larval and juvenile red drum in the Aransas Estuary varied as a function of both habitat (shoal grass Halodule wrightii, turtle grass Thalassia testudinum) and site (Aransas Bay, Redfish Bay); however, no differences in RNA:DNA ratios were detected between habitats or between sites. It is postulated that the nutritional condition of newly settled red drum from the Aransas Estuary in 1994 was relatively high, and that starvation was of minor importance. Received: 19 August 1996 / Accepted: 23 August 1996     

Effects of the polycyclic aromatic hydrocarbon fluoranthene on growth rate and nucleic acid composition of Capitella sp. I

Body volume growth rate, dry tissue weight, organic carbon content, and nucleic acid content (RNA and DNA) of individual Capitella sp. I were measured after 14 d of exposure to natural sediment, sediment contaminated with fluoranthene (in acetone) and sediment treated with acetone only. Exposure to 101 μg fluoranthene g⁻¹ sediment dry wt during 2 weeks reduced body volume specific growth rate by 50%. Dry tissue weight and carbon content were also lower in fluoranthene-exposed worms. However, when corrected for differences in body volume, worms from fluoranthene-contaminated sediment had a higher dry weight and carbon content per unit body volume than worms from the control and acetone treatments. Part, but not all, of the reduction in body volume growth rate in response to fluoranthene may have been due to a reduction in tissue water content. The correlation between RNA:DNA ratio (which has previously been used as a predictor of recent growth rate) and worm growth rate was weak in the control group and was significantly influenced by sediment treatment. Not only did the fluoranthene-exposed worms have a lower growth rate than expected based on their RNA:DNA ratio, but analysis of this group separately indicated that measured growth rate was independent of the RNA:DNA ratio. Our results not only confirm previous indications of a weak relationship between nucleic acid content and actual growth, but indicate that empirical, predictive relationships between these variables measured under favorable growth conditions should not be extrapolated to predict growth under toxicant-stressed conditions. Received: 6 April 1997 / Accepted: 7 May 1997