Responsiveness of selected condition measures of herring, Clupea harengus, larvae to starvation in relation to ontogeny and temperature

A laboratory experiment was performed to study the responsiveness of selected condition measures to starvation in herring, Clupea harengus, larvae in relation to temperature and ontogeny. The larvae at two intervals of development, i.e. stage 1 larvae with initial exogenous feeding and stage 2 larvae with established feeding prior to notochord flexion, were reared at three temperatures (5, 8 and 11°C) and subjected to sub-lethal durations of starvation. Temporal changes in standard length, dry weight, DNA concentration (% of dry weight), RNA concentration (% of dry weight), Fulton's condition factor (CF) and RNA/DNA were assessed and compared with fed controls. Starvation led to decreases in dry weight, CF, RNA concentration and RNA/DNA, while it led to an increase in DNA concentration. Higher responsiveness to starvation was observed at higher temperatures, and the magnitude of the changes was higher in stage 2 larvae. The shortest latency in starvation response was found with respect to RNA/DNA which was length independent in the size range studied. RNA/DNA was also significantly related to average DNA growth rate, and the model for DNA growth rate was, SGRDNA=4.49 RNA/DNA + 7.14 T – 0.42 T2 – 37.5 ; n=32, r2=0.85, p < 0.001). While the model seemed to adequately represent the average temperature dependent DNA growth, a relatively low classification success made it unsuitable for depicting individually starving larvae. Critical levels in DNA concentration can be used (2.2% for stage 1 larvae, 2.9% for stage 2 larvae) to differentiate starving larvae (after 3 – 5 days) from feeding larvae. RNA/DNA was the most sensitive and suitable condition index studied in detecting early starvation of herring larvae.     

Nutritional condition of larval milkfish,Chanos chanos,occurring in the surf zone

In order to clarify the nutritional conditions of larval milkfish in the surf zone, the following parameters were examined: 1) DNA and RNA content and RNA/DNA ratio of fed and unfed larvae collected from the surf zone and reared in the laboratory; 2) survival rate of the unfed larvae; and 3) total length, otolith increment counts and RNA/DNA ratio of wild larvae collected daily from the surf zone. The DNA and RNA content of the unfed larvae decreased, but increased in fed larvae. The RNA/DNA ratio decreased in unfed larvae, whereas in the fed larvae it decreased for the first three days after capture and increased thereafter. These results indicated that the values of DNA and RNA content and RNA/DNA ratio could be used as an indicator of nutritional condition of milkfish larvae after 6 days of starvation. Although total length of the wild-larvae did not show serial changes, their otolith increment counts showed continuous increases, indicating that the larvae sojourned in the surf zone for several days. In the same period, RNA/DNA ratios of the wild larvae decreased continuously, the ratios of larvae with fewer otolith increment counts being relatively higher than those of larvae with greater increment counts. Based on these results, the milkfish larvae remaining in the surf zone were concluded as being under insufficient nutritional conditions.     

Assessment of the nutritional status of field-caught larval Pacific bluefin tuna by RNA/DNA ratio based on a starvation experiment of hatchery-reared fish

RNA/DNA ratio is a useful and reliable indicator of the nutritional status of fish larvae and juveniles. In order to assess the nutritional status of field-caught larval Pacific bluefin tuna Thunnus orientalis (Temminck et Schlegel), starvation experiments of hatchery-reared larvae were conducted and changes in the RNA/DNA ratio of fed and starved larvae were analyzed. Starvation experiments were conducted every 3 days after first feeding. The survival rate of Pacific bluefin tuna larvae ranged 10-50% after 1 day of starved conditions and growth retardation was observed immediately. These results suggest that Pacific bluefin tuna larvae have a very low tolerance to starvation. The RNA/DNA ratios of fed larvae were approximately 2.0-4.0. On the other hand, the value of starved larvae significantly decreased to 1.0-3.0. The nutritional status of 3 cohorts of field-caught tuna larvae collected in the northwestern Pacific Ocean was examined based on the value of the RNA/DNA ratio of the 1 day starved larvae. 4.35-25.77% of the cohorts were regarded as the “starving condition”, which was negatively correlated to the ambient prey densities. These findings suggest that the nutritional condition of larval Pacific bluefin tuna was influenced by the ambient prey density, and starvation itself and starvation-induced predation could greatly contribute to mortality in the larval period of Pacific bluefin tuna.     

Nutritional condition and vertical distribution of Baltic cod larvae

Newly hatched Baltic cod Gadus morhua larvae are typically found at depths >60 m. This is a region of low light and prey availability, hence generating the hypothesis that larvae have to migrate from hatching depth to the surface layer to avoid starvation and improve their nutritional condition. To test this hypothesis, Baltic cod larvae were sampled during the spawning seasons of 1994 and 1995 with depth-resolving multiple opening/closing nets. Each larva was aged by otolith readings and its RNA/DNA ratio was determined as a measure of nutritional condition. The RNA/DNA ratios of these larvae aged 2-25 days (median 10 days) ranged from 0.4 to 6.2, corresponding to levels exhibited by starving and fast-growing larvae in laboratory calibration studies (starvation, protein growth rate, G_pi= -12.2% day⁻¹; fastgrowing larvae, G_pi=14.1%day⁻¹) respectively. Seventy per cent of the field caught larvae had RNA/DNA ratios between the mean values found for starving and fed laboratory larvae. Only larvae aged 8-11 days had higher mean RNA/DNA ratios above 45 m than below ( t -test, P<0.05). However, the instantaneous protein growth rates were significantly higher for all larval age groups in the surface layers ( t -test, P<0.05). Starving larvae were found in all depths sampled (10-85 m), whereas growing larvae (positive G_pi) were restricted to samples taken shallower than 45 m. These superior growth rates above 45 m corroborate the hypothesis and imply that migration to the shallow water layers is a prerequisite for good nutritional condition, growth and survival of Baltic cod larvae. The frequent occurrence of cod larvae older than 8 days in the deep water in poor condition suggests that a proportion of the larvae will die from Starvation in the deep layers of the Baltic Sea.     

Nutritional condition, starvation status and growth of early juvenile Japanese sea bass (Lateolabrax japonicus) related to prey distribution and feeding in the nursery ground

The nutritional condition and protein growth rates of Japanese temperate bass larvae and juveniles were studied in relation to prey distribution and feeding habits in the nursery grounds in Chikugo estuary, Ariake Sea, Japan. Samples were collected from a wide spatial area covering the nursery grounds of the fish in March and April 2003. Food habits of the fish were analyzed by examining the gut contents. Fish condition was evaluated by using RNA/DNA ratio and other nucleic acid-based indices and protein growth rates. The nucleic acid contents in individually frozen larvae and juveniles were quantified by fluorometric method. Two distinguished feeding patterns, determined by the distribution of prey copepods, were identified along the nursery ground. The first pattern showed the dependency of the fish on the calanoid copepod Sinocalanus sinensis, which was the single dominant prey in low saline upper river areas and the second pattern involved a multi-species dietary habit mainly dominated by Acartia omorii, Oithona davisae and Paracalanus parvus. Values of RNA, DNA, total protein, growth rates and for all the nucleic acid-based indices were higher in upstream areas than in the downstream areas. The proportion of the starving fish was higher in the downstream areas than in the upstream areas. Condition of juvenile sea bass was not equal throughout the nursery grounds; fish in the upper river were in better condition than those in the lower estuary. We speculated that utilization of S. sinensis, which appears a suitable prey item and provide a better foraging environment in the upstream nursery ground, is one of the key factors for early survival and growth of Japanese temperate bass larvae and juveniles in the Chikugo estuary.     

Feeding, Ichthyophonus sp. infection, distribution and growth history of Norwegian spring-spawning herring in summer

After its collapse in the late 1960s, 1991 was the first year of systematic herring investigations after the Norwegian Sea stock returned to the traditional summer-feeding areas. Of 11 prey groups three dominated the diet; Calanus finmarchicus, Parathemisto spp. and Sebastes spp. There was a high prevalence of Ichthyophonus sp. at stations in the central and western parts of the Norwegian Sea where the cpue was low. This suggests that infected individuals were unable to return from the feeding to the wintering area of the Norwegian fjords. Highest cpue were recorded between 68–73° N and 5–15° E. Age groups 2, 3 and 8 were most frequently caught and age group 8 (the 1983 year class) dominated the samples. Herring occurred in dense concentrations where C. finmarchicus dominated diet. 0-group fish dominated diet at stations along the coast from 66–73° N and out to c . 300 km off the coast. Mature herring overlapped in distribution with 0-group fish and Sebastes spp. contributed largely to herring diet in this area. The first rich year-class after the collapse, the 1983 year class, had a faster growth than rich year classes from past herring periods, but slower than poor ones.     

A model to estimate growth in young-of-the-year tautog, Tautoga onitis, based on RNA/DNA ratio and seawater temperature

This study presents an indirect method for estimating growth rates of young-of-the-year (YOY) tautog, Tautoga onitis, based on laboratory calibration experiments and nucleic acid-based indices. Field-collected tautog were held in the laboratory at 3 temperatures over a 17-day period. Four feeding levels were used to produce a range of growth rates. An ultraviolet absorption assay was used to measure nucleic acid concentrations in white muscle tissue. The strength of the relationship between growth rate and three nucleic acid-based parameters (RNA concentration, DNA concentration, RNA/DNA ratio (R/D)) was tested. Correlation results indicated a significant positive relationship between R/D and weight-based instantaneous growth rate (G) (r = 0.68; p < 0.001). Both R/D (r = − 0.55; p < 0.006) and RNA (r = − 0.56; p < 0.005) were highly negatively correlated with temperature (T). Multiple linear regression showed that R/D and temperature explained 61% of the variability in growth, resulting in the model G = 0.01285(R/D) + 0.00057(T) − 0.03205 (p < 0.0001). This R/D-temperature model can be used to evaluate recent growth rates in YOY tautog under field conditions and has applications for aquaculture when comparing growth rates of fish held under different culture conditions.     

Evaluating the nutritional condition of individual whitefish (Coregonus spp.) larvae by the RNA/DNA ratio

Larvae of Coregonus spp. were reared in the laboratory under different temperature (4, 6 and 8°C) and feeding conditions ( ad libitum , limited, with starvation intervals). Their RNA/DNA ratios were determined with a highly sensitive fluorescence technique. After resorption of their yolk reserves (about 2 weeks after hatching), well fed larvae (RNA/DNA >2.5) could be significantly distinguished from larvae reared under limited food supply (RNA/DNA < 2.5), both at the 6 and 8°C levels. At 4°C, no differences due to the feeding regime were found. During a second series of experiments, larvae were affected by an intestinal disease, which was caused by the ingestion of unsuitable copepod plankton. This disease provoked high mortality, decreased growth and RNA/DNA ratios which were almost as low as in temporarily starved larvae from the first series. Coregonid larvae sampled in Lake Constance during spring 1990 showed RNA/DNA ratios which were unexpectedly low when judged on the basis of mean body length and average ambient temperature. It was obvious from macroscopic observations that some of these wild larvae were severely attacked by the intestinal disease. The low RNA/DNA ratios in field samples are, therefore, interpreted as a sublethal result of this disease.     

Analysis of covariance: an alternative to nutritional indices

Some statistical problems are added to the growing list of cautionary tales regarding the use of the conventional, ratio-based nutritional indices (RCR, RGR, ECI, AD and ECD). Analysis of ratios is based on the, probably unrealistic, assumption of an isometric relationship between denominator and numerator variables. Analysis of covariance (ANCOVA) makes less restrictive assumptions, and additionally provides important information about the data which is lost by using ratio variables. We demonstrate, using computer-generated data sets, some of the pitfalls of statistical analysis of ratios and illustrate how these may be avoided using ANCOVA. Some possible consequences of such statistical iniquities for biological interpretations are discussed.     

Field evaluation by RNA/DNA ratios on post-release nutritional status of released and wild Japanese flounder Paralichthys olivaceus juveniles

A large-scale juvenile Japanese flounder (Paralichthys olivaceus) release-recapture experiment was undertaken to find the optimal release season by evaluating the nutritional status of released fish at different seasons during which food abundance was significantly different. Forty thousand fish were released at depths of 1.5 m for early-release (May 29, 1997) and 2 m for late-release (July 2, 1997) (42.1±3.5 and 40.9±4.2 mm body length, respectively) in an experimental field, Wakasa Bay, the Sea of Japan. Samples were taken, after the releases, at Wada beach at intervals of 1, 2, 3, 6, 10, 16 and 30 days after release (DAR), including pre-surveys before each release. Released fish recaptured from the two different release groups totaled 764; 467 from the early-release group (ER) and 297 from the late-release group (LR). A total of 1956 wild flounder juveniles were simultaneously collected (1041 ER, 915 LR). ER fish were subject to higher food availability and were exposed to less pressure from predation by smaller wild juvenile flounder. RNA/DNA ratios in ER juveniles were significantly higher than those of LR fish during all samples. Especially, RNA/DNA ratios in ER juveniles were higher than in wild juveniles from 3 to 50 DAR. In the LR group, the nutritional status of juveniles was relatively low in shallower water. These findings corresponded well with feeding incidence examined by coworkers. Mass release of hatchery-reared juveniles apparently reduced RNA/DNA ratio of the wild juveniles right after releasing. The present study showed that earlier release of hatchery-reared juvenile Japanese flounder with higher RNA/DNA ratio could increase the possibilities of survival right after release in the nursery ground, and that RNA/DNA ratio appeared to be a good tool in evaluating nutritional status of released juveniles as well as wild juveniles in Japanese flounder.     

Sources of variation for nutritional condition indices of the plasma of migratory lesser kestrels in the breeding grounds

Although published information on reference values for biochemical parameters in birds of prey has increased during the last years, little is known on their sources of variation. We used an insectivorous and small migratory raptor species, the lesser kestrel Falco naumanni, as a model. We looked for sources of variation of nutritional biochemical parameters (i.e. triglycerides, cholesterol, uric acid and urea) of both nestlings and adults. Reference values indicated that, as a rule, lesser kestrel showed more elevated triglycerides, urea and uric acid levels than other raptors. All analyzed factors except gender (i.e. year, colony, sampling time, presence/absence of a geolocator, body mass, laying date and capture date) reached significance for at least one biochemical parameter. In the morning, we found an important postprandial increase in the concentration of all biochemical parameters in nestlings, and uric acid and urea levels in adults. A positive relationship was detected between triglycerides and body mass in nestlings and adults. Although we did not find differences between blood biochemical parameters of the oldest and youngest chick of each brood, we found that cholesterol levels were lower in nestlings from larger broods. Coloration of tarsi (measured as brightness) was related to triglycerides and urea levels of nestlings and adults, respectively. The feeding habits of lesser kestrel probably explain the different levels and patterns of variation of metabolites in comparison to more carnivorous raptors eating mammals or birds.     

Strontium:cakium concentration ratios in otoliths of herring larvae as indicators of environmental histories

Synopsis Elemental analyses, using wave-length dispersive electron microprobe techniques on otoliths from reared Atlantic herring larvae, Clupea harengus, showed trace quantities of strontium relative to that of calcium, and an inverse relationship between Sr/Ca concentration ratios and rearing temperature. These data are consistent with those for coral aragonite, in that there appears to be an inverse temperature effect on physiological incorporation of strontium in the otolith aragonite. Our determinations of Sr/Ca concentration ratios of lab-reared herring larvae showed that the deposition of strontium relative to calcium and the rearing temperature were related, where: T (° C) = −2.955 [Sr/Ca] × 1000 ± 19.172. This principle thus makes it possible to use Sr/Ca concentration ratios in fish otoliths to delineate past temperatures experienced by an individual. Further, combining electron microprobe analyses with scanning electron microscope (SEM) examinations of daily increments in the same otolith makes it possible to reconstruct the temperature history for an individual fish on a time scale of days. An example of the application of the technique to an approximately six-month-old field-caught herring larva is given, and the limitations of the technique are discussed.     

In vitro reconstitution and characterization of the yeast mitochondrial degradosome complex unravels tight functional interdependence

The mitochondrial degradosome (mtEXO), the main RNA-degrading complex of yeast mitochondria, is composed of two subunits: an exoribonuclease encoded by the DSS1 gene and an RNA helicase encoded by the SUV3 gene. We expressed both subunits of the yeast mitochondrial degradosome in Escherichia coli, reconstituted the complex in vitro and analyzed the RNase, ATPase and helicase activities of the two subunits separately and in complex. The results reveal a very strong functional interdependence. For every enzymatic activity, we observed significant changes when the relevant protein was present in the complex, compared to the activity measured for the protein alone. The ATPase activity of Suv3p is stimulated by RNA and its background activity in the absence of RNA is reduced greatly when the protein is in the complex with Dss1p. The Suv3 protein alone does not display RNA-unwinding activity and the 3' to 5' directional helicase activity requiring a free 3' single-stranded substrate becomes apparent only when Suv3p is in complex with Dss1p. The Dss1 protein alone does have some basal exoribonuclease activity, which is not ATP-dependent, but in the presence of Suv3p the activity of the entire complex is enhanced greatly and is entirely ATP-dependent, with no residual activity observed in the absence of ATP. Such absolute ATP-dependence is unique among known exoribonuclease complexes. On the basis of these results, we propose a model in which the Suv3p RNA helicase acts as a molecular motor feeding the substrate to the catalytic centre of the RNase subunit.     

Climate change and condition of herring (Clupea harengus) explain long-term trends in extent of skipped reproduction

It is commonly assumed that iteroparous fish, once mature, normally reproduce in all consecutive seasons. Recent work has suggested, however, that in Norwegian spring-spawning herring—a population that undertakes extensive spawning migrations—almost one in two adults may skip their second spawning migration. Why should herring not return to spawn the year after first spawning, but instead wait an extra year? For herring, participation in distant, energetically costly, and risky spawning migrations will only pay off in terms of fitness if individuals are sufficiently large, and in sufficient condition, to both successfully migrate and spawn. Changes in the environment and individual condition should therefore affect the likelihood of skipped spawning. This paper describes long-term changes in the extent to which the second reproductive season is skipped in this herring population. These are shown to be linked to the size and condition of herring as first-time spawners, and to climatic factors possibly related to food availability. The findings corroborate the hypothesis that skipped reproduction results from trade-offs between current and future reproduction, growth and survival.     

Method of predicting reference condition biota affects the performance and interpretation of ecological indices

1. The statistical rigour and interpretability of ecological assessments is strongly affected by how well we predict the biological assemblages expected to occur in the absence of human‐caused stress, i.e. the reference condition. In this study, we examined how the specific method used to predict the reference condition affected the performance of two commonly used types of ecological index: RIVPACS‐based O/E indices and multimetric indices (MMIs). 2. These two types of index have generally relied on different approaches to predicting the reference condition. For MMIs, some type of regionalisation is typically used to describe the range of metric values among reference sites and hence the expected range at assessed sites. For O/E indices, continuous modelling is used to predict how the biota varies among sites both among and within regions. Because the prediction method differs for these two types of index, it has been impossible to judge if differences in index performance (accuracy, precision, responsiveness and sensitivity) are caused by differences in the way reference condition biota are predicted or by differences in what the indices measure. 3. We used a common data set of 94 reference sites and 255 managed sites and the same potential set of predictor variables to compare the performance of five different MMIs and three O/E indices that were derived from different prediction methods: null models, multiple linear regression (MLR), classification and regression trees, Random Forests (RF) and linear discriminant functions models (LDM). We then calculated values of these indices for samples collected from the managed catchments as well as samples collected from 13 reference sites that were progressively altered in known ways by a simulation programme. 4. Both the type of predictor used and the type of index affected overall index performance. Modelled indices generally had the greatest sensitivity in assessing managed sites as biologically different from reference. Index sensitivity was determined by both an aspect of index precision (10th percentile of reference condition values) and responsiveness. The O/E indices showed the best scope of response to known biological alteration. All three O/E indices decreased linearly in response to simulated alteration in both overall assemblage structure (Bray‐Curtis dissimilarity) and taxa loss. The MMIs declined linearly from low to intermediate levels of assemblage alteration but were less responsive between intermediate and high levels of biological alteration. 5. Insights gained from simulations can aid in testing assumptions regarding index response to stress and help ensure that we select indices that are ecologically interpretable and most useful to resource managers.     

Nucleic acids in ichthyoplankton ecology: a review, with emphasis on recent advances for new perspectives

Nucleic acid assessments and especially the RNA/DNA ratio are used widely as indices of growth rate or nutritional condition of larval fish. Methodological aspects are considered with the aim of defining a most reliable analytical procedure. Laboratory calibrations can give conflicting results, but general and common features may be drawn from a global examination. A high variability of RNA content, and thus RNA/DNA ratio, was revealed by estimates at the individual level and seems to be more related to feeding condition in late larval or juvenile stages than in yolk-sac and first-feeding larvae. An alternative is suggested, based on the relative DNA content which appears more stable and sensitive to starvation during these early stages. The consistency of such a pattern is strongly supported by findings about regulation of ribosomal RNA content of tissues, and especially white muscle, by nutrition. Determination of a threshold founded on a simple observation of morphological development stages is proposed as a base to define the best way to assess the larval nutritional condition. Some field applications are reviewed briefly. Finally an appraisal of the advantages and drawbacks of the different nucleic acid-based indices is attempted, which permits delineation of some prospects for future research.