Feeding energetics and metabolism in demersal fish species from Antarctic,temperate and tropical environments

The energetics of feeding has been investigated in demersal fish with similar sedentary lifestyles from the Antarctic (Notothenia neglecta Nybelin), North Sea (Myoxocephalus scorpius L.) and Indian Ocean (Cirrhitichys bleekeri Bleeker). In general, the metabolic rates of fasting individuals were positively correlated with adaptation temperature: values for a standard 100 g fish (mg O₂/h) were 3.3 for N. neglecta at around 0 °C, 2.7 for winter-acclimatized M. scorpius at 5 °C, 4.3 for summer-acclimatized M. scorpius at 15 °C, and 7.0 for C. bleekeri at 25 °C. In all species, following a single satiating meal, oxygen consumption increased to a peak of 2 to 3.5 times the fasting values. Maximum rates of oxygen consumption after feeding were several-fold higher in the warm-than in the cold-water species. After controlling for the effects of body mass and energy intake by analysis of covariance, the duration of the increase in metabolic rate, referred to as specific dynamic action (SDA), was found to be 3 to 4 times shorter in the warm- than in the cold-water fish, ranging from 57 h in C. bleekeri to 208 h in N. neglecta. In contrast, the SDA was not significantly different in the various species, corresponding to 15 to 23% of the energy ingested. Seasonal influences on metabolism and feeding were also studied in N. neglecta acclimated to simulated winter (-1.0 to-0.5 °C; 3 h light:21 h dark) or summer (0 to 0.9 °C; 21 h light:3 h dark) conditions. The metabolic rates of fasting and fed individuals, and the characteristics of the SDA were found to be independent of acclimation conditions. This suggests that N. neglecta is capable of processing food at similar rates throughout the year. Energy stores and enzyme activities were measured in the swimming muscles and liver of fish fed ad libitum. Summer-acclimated fish had higher concentrations of liver triglyceride stores and elevated activities of some enzymes of intermediary metabolism relative to winter-acclimated fish. The observed changes in intermdiary metabolism are probably related to annual cycles of growth and reproduction. It is suggested that the low aerobic scope for physiological performance in Antarctic fish may necessitate the seasonal switching of energy allocation between growth and reproduction.     

Locomotory activity and behaviour of the Antarctic teleostNotothenia coriiceps

The activity and behaviour of a free-living Antarctic fish,Notothenia coriiceps Richardson (formerlyN. neglecta), was investigated using a high-sensitivity, underwater TV camera at Signy Island, South Orkney Islands. Detailed observations of the 33 cm TL (total length) fish were made over a period of 6 d in austral summer (February 1992), for a total 69.5 h. Natural light at 2.5 m depth allowed viewing from 1 h before sunrise to 1 h after sunset. The fish stayed in a territory within 3 m of a small cave for >98% of the time, and made between 1 to 148 swims d^-1, of which 92.5% were brief (<15 s) feeding attempts. On average, 1.7% of each day was engaged in locomotion, including 1.2% swimming and 0.5% manoeuvring. Swimming was generally slow, at <2 body lengths s^-1, and labriform and subcarangiform modes were used alternately or in combination. Activity level (swims or displays per hour) was unaffected by tide, but was lower for 3 d when a wind speed >16 knots prevailed indicating that large waves reduced activity. A suspected diurnal activity rhythm was not statistically significant. The fish is an ambush-predator, and it took most of its prey from the water column but some off macroalgae or the seabed. Ventilation rate was slightly higher after activity, and peaked after an encounter with anotherN. coriiceps.     

Biochemical adaptations of the mudskipper Boleophthalmus boddaerti to a lack of oxygen

Specimens of the mudskipper Boleophthalmus boddaerti, collected along the estuarine canal at Pasir Ris, Singapore between April 1987 and December 1989, were examined in the laboratory. After being exposed to environmental hypoxia for 6 h, no accumulation of lactate, alanine or succinate was observed in the muscle tissue of B. boddaerti. In addition, the blood lactate content and muscle creatine phosphate (CrP), adenosine triphosphate (ATP) and glycogen contents were not significantly different from those of the control. During normoxic recovery, oxygen debt repayment was only a small fraction of the oxygen deficit incurred during the 6 h of hypoxic exposure. Therefore, it would appear that B. boddaerti coped with environmental hypoxia by undergoing metabolic rate reduction. After anoxic exposure, the CrP, ATP and adenosine diphosphate (ADP) contents in the muscle tissue of B. boddaerti were significantly lower than the corresponding control values. Lactate accumulated in the blood and muscle tissue of the anoxic fish though the muscle energy charge and glycogen content remained constant. Hence, B. boddaerti was able to cope with such a period of anoxia without increasing its glycolytic rate. Succinate, which was not detected in the muscle tissue of the control and hypoxia-exposed fish, accumulated in significant quantities in the muscle tissue of B. boddaerti exposed to environmental anoxia and functional hypoxia. In the case of functional hypoxia, there was a significant decrease in the muscle glycogen content, and the muscle lactate content increased 17.3-fold, indicating that glycolysis was activated to provide the source of energy during muscular exercise.     

Metabolic scaling,buoyancy, and growth in larval Atlantic menhaden,Brevoortia tyrannus

We measured growth of larval Atlantic menhaden, Brevoortia tyrannus in terms of mass, volume, and weight in water as well as the mass-specific activities of the metabolic enzymes citrate synthase, lactate dehydrogenase, and malate dehydrogenase. Weight in water, the force the fish must exert to maintain vertical position, increases by a factor of 18 in larvae growing from 10 to 15 mm. The weight increase coincides with the development of the larval swim bladder. The activities of all three enzymes per unit mass of fish tissue decline greatly over this time period, indicating that the fish does not further develop its aerobic and anaerobic metabolic capacity for swimming during this growth interval.     

Niche overlap in mud snails (hydrobiidae): freezing tolerance

Tolerance to freezing was measured in four species of hydrobiid snails exposed to temperatures from 0° to-8°C and salinities from 0 to 30 S four up to 7 d. Experiments showed the following increasing order of tolerance to freezing: Potamopyrgus jenkinsi (Smith), Hydrobia neglecta Muus, H. ulvae (Pennant) and H. ventrosa (Montagu). Survival decreased with increasing salinity in P. jenkinsi, while the opposite was found in Hydrobia species. H. ulvae survived only slightly better than H. neglecta, while H. ventrosa was very tolerant to freezing even at low salinities in contrast to the other species of Hydrobia. H. neglecta suffered higher winter mortality than H. ventrosa during the severe winter of 1985 in an estuary.     

Sperm swimming speed and energetics vary with sperm competition risk in bluegill (<Emphasis Type="Italic">Lepomis macrochirus</Emphasis>)

Under sperm competition, a males fertilization success depends largely on the ejaculate characteristics of competing males. Theoretical models predict that, in external fertilizers, increased risk of sperm competition should result in selection for increased sperm swimming speed. To test this prediction, we studied the behavior of sperm from parental and sneaker male bluegill (Lepomis macrochirus), a fish species characterized by high levels of cuckoldry due to alternative reproductive tactics of males (parentals and cuckolders). Because cuckolders (sneakers and satellites) always spawn in the presence of a parental male, but the reverse is not true, cuckolders experience the greater risk of sperm competition. We show here that the spermatozoa of sneakers have faster initial swimming speeds but shorter periods of motility than the sperm of parental males. Moreover, we show that sperm swimming speeds shortly after activation (when most fertilization occurs) are correlated with starting ATP levels in spermatozoa, suggesting that sperm competition has selected for higher energetic capacity in the sperm of sneakers. Thus, the higher energetic capacity and initial swimming speed of sneaker sperm may explain why, despite having fewer sperm per ejaculate than parentals, sneakers fertilize more eggs than parental males when they compete to fertilize a clutch of eggs.Communicated by L.W. Simmons     

External devices on penguins: how important is shape?

Many researchers use external recording or transmitting devices to elucidate the marine ecology of fish, mammals and birds. Deleterious effects of these instruments on the parameters researchers wish to measure are hardly ever discussed in the literature. Research has shown that, in penguins, volume and cross-sectional area of instruments negatively correlate with swimming speed. dive depth and breeding success, and that device colour affects bird behaviour. Here, a large (200 g, cross-sectional area 2100 mm²) streamlined device was attached to the lower back of Adélie penguins (Pygoscelis adeliae on Ardley Island, South Shetland Island in 1992) and its effects on bird swimming speed and energetics were measured in a water canal in Antarctica. Although the device was 10.5% of penguin cross-sectional area, swimming speed was reduced by only 8.3% and mean power input increased by only 5.6% while swimming. Although our streamlined device was five times more voluminous than one of our older units, the effect on swimming energetics could be reduced by 87%.     

Right tools for the task: intraspecific modality in the swimming behaviour of coral reef fishes

Understanding the movement patterns of fishes in the wild requires an awareness of the intrinsic and extrinsic factors underlying their behaviour. Using field observations, we explored whether eight species of coral reef fish display modality in their swimming behaviour, which is a convergence in fin use patterns among and within species when they conduct daily activities (e.g. feeding, travelling, chasing). While a range of different fins were used during steady swimming by each species, most fishes converged towards median-paired fin use (e.g. pelvics, pectorals) when undertaking searching and feeding. Conversely, all species adopted caudal fin use during high-speed activities such as chasing. Such modality in fin use was relatively conserved across juvenile and adult conspecifics and transcended differences in phylogenetic history and trophic preferences, suggesting that overarching constraints underpin patterns of fish swimming behaviour in complex habitats.     

Microplastic contamination in Great Lakes fish

Freshwater ecosystems, generally adjacent to human population and more contaminated relative to adjacent marine ecosystems, are vulnerable to microplastic contamination. We sampled 7 species of fish from Lake Ontario and Lake Superior and assessed their gastrointestinal (GI) tracts to quantify ingested microplastics and other anthropogenic particles. A subset of the microparticles were chemically analyzed to confirm polymer types and anthropogenic origins. We documented the highest concentration of microplastics and other anthropogenic microparticles ever reported in bony fish. We found 12,442 anthropogenic microparticles across 212 fish (8 species) from nearshore Lake Ontario, 943 across 50 fish (1 species) from Humber River, and 3094 across 119 fish (7 species) from Lake Superior. Fish from Lake Ontario had the greatest mean abundance of anthropogenic microparticles in their GI tracts (59 particles/fish [SD 104]), with up to 915 microparticles in a single fish. Fish from Lake Superior contained a mean [SD] of 26 [74] particles/fish, and fish from Humber River contained 19 [14] particles/fish. Most particles were microfibers. Overall, ≥90% of particles were anthropogenic, of which 35-59% were microplastics. Polyethylene (24%), polyethylene terephthalate (20%), and polypropylene (18%) were the most common microplastics. Ingestion of anthropogenic particles was significantly different among species within Lake Ontario (p < 0.05), and the abundance of anthropogenic particles increased as fish length increased in Lake Ontario (ρ = 0.62). Although we cannot extrapolate the concentration of microplastics in the water and sediments of these fish, the relatively high abundance of microplastics in the GI tracts of fish suggests environmental exposure may be above threshold concentrations for risk.     

Distribution of enzymes of glycolysis and gluconeogenesis in fish tissues

Gluconeogenesis in fishes has been demonstrated in whole animals and liver preparations. However, at present, the relative physiological importance of possible substrates such as lactate, pyruvate and amino-acids or the precise sites of gluconeogenesis are unclear. In mammals, gluconeogenesis takes place in the liver and kidney, and the same could occur in fishes although it has been proposed that fish red muscle is also capable of reconverting lactate (derived from white muscle) to glucose. In this present study, the activities of 3 key glycolytic (hexokinase, phosphofructokinase and pyruvate kinase) and 2 key gluconeogenic (fructose diphosphatase and phosphoenolpyruvate carboxykinase) enzymes were investigated in tissues of the rainbow trout Salmo gairdneri, the cod Gadus morhua, and the plaice Pleuronectes platessa in order to elucidate the relative glycolytic/gluconeogenic capacities of the individual fish tissues. The glycolytic enzymes were found in all tissues, the relative potential being skeletal muscle>heart, brain >kidney, gills>liver. The gluconeogenic enzymes were not present in all tissues, and were mainly concentrated in the liver and kidney. Hence the results indicate that the liver, and to a lesser degree, the kidney are the major sites of gluconeogenesis in fishes, and that the process is unlikely to occur in skeletal muscle.     

Propulsion efficiency and cost of transport for copepods: a hydromechanical model of crustacean swimming

In the absence of direct measurement, costs of locomotion to small swimming Crustacea (<10 mm) have been derived exclusively through application of the fluid dynamic theory. Results indicate very low swimming costs, and contradict experimental data on larger Crustacea (15 to 100 mm) that suggest a three-fold increase in metabolic rate with increasing swimming speed. This paper introduces a swimming model that analyzes the hydrodynamic forces acting on a crustacean swimming at non-steady velocity. The model treats separately the hydrodynamic forces acting on the body and the swimming appendages, approximating the simultaneous solution of equations quantifying the drag and added-mass forces on each by stepwise integration. Input to the model is a time-series of instantaneous swimming-appendage velocities. The model output predicts a corresponding time-series of body velocities as well as the mechanical energy required to move the swimming appendages, dissipated kinetic energy, and metabolic cost of swimming. Swimming of the calanoid copepod Pleuromamma xiphias (Calanoida) was analyzed by extrapolating model parameters from data available in the literature. The model predictions agree well with empirical observations reported for larger crustaceans, in that swimming for copepods is relatively costly. The ratio of active to standard metabolism for P. xiphias was >3. Net cost of transport was intermediate to the values found experimentally for fish and larger crustaceans. This was a consequence of the predicted mechanical efficiency (34%) of the copepod's paddle propulsion, and of increased parasitic resistance resulting from non-steady velocity swimming.     

Predator shoaling moderates the confusion effect in blue-green chromis,Chromis viridis

Summary In experiments, blue-green chromis [Chromis viridis (Cuvier 1830)] were fed on either scattered or aggregated swarms of brine shrimp (Artemia sp.). Ten runs with each prey dispersion treatment were performed with shoals of one, two, five and ten chromis. The mean lag in reaching peak feeding rate for fish fed on aggregated prey was significantly shorter in the larger chromis shoals. In contrast, with the scattered treatment all such lags were similar and very short. As foraging proceeded, higher feeding rates were observed in the larger feeding shoals, regardless of prey dispersion. Prey capture success (i.e. the rate of retention of intercepted prey) declined with time, but was significantly higher in groups of ten fish. Two main conclusions emerge. Firstly, grouping facilitated initiation of feeding by individuals preying on concentrated swarms and reduced the delay in reaching a maximum feeding level. This may have been due to a suppression of the confusion effect through reduced reliance upon vigilance. Secondly, reduced vigilance allowed larger shoals of chromis to feed effectively over more extended periods. Trends of increasing shoal cohesion and decreasing prey retention rate with time were consistent with a postulated increase in antipredator vigilance with declining feeding motivation.     

Oxidative stress and genotoxicity of 1-methyl-3-octylimidazolium chloride on loach (Misgurnus anguillicaudatus)

This study was a preliminary step to evaluate the acute toxicity of 1-methyl-3-octylimidazolium chloride ([C₈mim]Cl) on loach (Misgurnus anguillicaudatus) by determining the effects on hepatic antioxidant enzyme activities and by the comet assay. The results showed that [C₈mim]Cl had acute toxicity at concentrations above 20 mg L^?1, inducing oxidative stress and genotoxicity on fish liver cells. In respect to enzyme activities, [C₈mim]Cl induced changes in the activities of superoxide dismutase, catalase, and glutathione content the livers of fish exposed at 20–80 mg L^?1. [C₈mim]Cl at the same exposure level caused a remarkable increase in malondialdehyde level. The comet assay indicated that [C₈mim]Cl at 20–80 mg L^?1 induced genotoxicity in liver cells. With increased exposure concentration and time, the two comet parameters trailing rate and tail moment were significantly increased, with significant differences (P < 0.05) observed between control group and each treatment group. The present study shows that ionic liquids can be a threat to the health of aquatic organism when accidentally released to aquatic ecosystems.     

Na+-K+-adenosine triphosphatase activities in gills of marine teleost fishes: Changes with depth,size and locomotory activity level

Activities of the primary enzyme responsible for monovalent ion regulation, Na⁺-K⁺-adenosine triphosphatase (Na⁺-K⁺-ATPase), were measured in gills of marine teleost fishes with different depths of occurrence (0 to 4800 m), body weights (a range of five orders of magnitude), and locomotory capacities. Specimens were collected off the coasts of California and Oregon in 1983–1989, and at the Galápagos Spreading Center and 13°N East Pacific Rise hydrothermal vent sites in 1987 and 1988, respectively. Except for two hydrothermal vent fishes, deep-sea species had much lower Na⁺-K⁺-ATPase activities g^–1 gill filament than shallow-living species, indicating that osmoregulatory costs, like total metabolic rate, are greatly reduced in most deep-living fishes. Within a species, the total branchial Na⁺-K⁺-ATPase activity per individual was dependent on size; the average allometric scaling exponent was 0.83. Using published values for oxygen consumption rates, and the total branchial Na⁺-K⁺-ATPase activities as an index of osmoregulatory costs, we estimated the maximal cost (as percent of ATP turnover) for osmoregulation in ten teleosts. Osmoregulatory costs averaged about 10% of total ATP turnover among these species, and maximal costs were no greater than about 20%. The percent costs of osmoregulation did not differ between shallow- and deep-living fishes. The reduced total ATP expenditure for osmoregulation in deep-living fishes is proposed to result from the sluggish locomotory habits of these fishes, not from selection for reduced osmotic coastper se. Thus, the reduced swimming abilities of these fishes lead to lower rates of water flow over the gills and less blood flow through the gills due to reduced demands for oxygen. Consequently, passive flux of water and ions through the gills is much lower than in more active fishes, and osmotic costs are thereby minimized. The relatively high activities of Na⁺-K⁺-ATPase in gills of the two hydrothermal vent fishes suggest that these fishes may be more active and have higher metabolic rates than other deep-sea fishes.     

Tail beat frequency as a predictor of swimming speed and oxygen consumption of saithe (<Emphasis Type="Italic">Pollachius virens</Emphasis>) and whiting (<Emphasis Type="Italic">Merlangius merlangus</Emphasis>) during forced swimming

Oxygen consumption and tail beat frequency were measured on saithe (Pollachius virens) and whiting (Merlangius merlangus) during steady swimming. Oxygen consumption increased exponentially with swimming speed, and the relationship was described by a power function. The extrapolated standard metabolic rates (SMR) were similar for saithe and whiting, whereas the active metabolic rate (AMR) was twice as high for saithe. The higher AMR resulted in a higher scope for activity in accordance with the higher critical swimming speed (U _crit) achieved by saithe. The optimum swimming speed (U _opt) was 1.4 BL s⁻¹ for saithe and 1.0 BL s⁻¹ for whiting with a corresponding cost of transport (COT) of 0.14 and 0.15 J N⁻¹ m⁻¹. Tail beat frequency correlated strongly with swimming speed as well as with oxygen consumption. In contrast to swimming speed and oxygen consumption, measurement of tail beat frequency on individual free-ranging fish is relatively uncomplicated. Tail beat frequency may therefore serve as a predictor of swimming speed and oxygen consumption of saithe and whiting in the field.     

The effects of zooplankton swimming behavior on prey-capture kinematics of red drum larvae, <Emphasis Type="Italic">Sciaenops ocellatus</Emphasis>

Most marine fishes undergo a pelagic larval phase, the early life history stage that is often associated with a high rate of mortality due to starvation and predation. We present the first study that examines the effects of prey swimming behavior on prey-capture kinematics in marine fish larvae. Using a digital high-speed video camera, we recorded the swimming velocity of zooplankton prey (Artemia franciscana, Brachionus rotundiformis, a ciliate species, and two species of copepods) and the feeding behavior of red drum (Sciaenops ocellatus) larvae. From the video recordings we measured: (1) zooplankton swimming velocity in the absence of a red drum larva; (2) zooplankton swimming velocity in the presence of a red drum larva; and (3) the excursion and timing of key kinematic events during prey capture in red drum larvae. Two-way ANOVA revealed that: (1) swimming velocity varied among zooplankton prey; and (2) all zooplankton prey, except rotifers and ciliates, increased their swimming velocity in the presence of a red drum larva. The kinematics of prey capture differed between two developmental stages in S. ocellatus larvae. Hyoid-stage larvae (3–14 days old) fed on slow swimming B. rotundiformis (rotifers) while hyoid-opercular stage larvae (15 days and older) ate fast moving A. franciscana. Hyoid-opercular stage red drum larvae had a larger gape, hyoid depression and lower jaw angle, and a longer gape cycle duration relative to their hyoid-stage conspecifics. Interestingly, the feeding repertoire within either stage of red drum development was not affected by prey type. Knowledge of the direct relationship between fish larvae and their prey aids in our understanding of optimal foraging strategies and of the sources of mortality in marine fish larvae.     

The effect of acute hypoxia on swimming stamina at optimal swimming speed in flathead grey mullet <Emphasis Type="Italic">Mugil cephalus</Emphasis>

Flathead grey mullets Mugil cephalus are commonly found in Mediterranean lagoons, which are regularly subject to high environmental variations. Oxygen is one of the factors that shows extremely high variation. The objective of this study was to test the effects of acute hypoxia exposure at two experimental temperatures (i.e. 20 and 30°C) on the stamina (time to fatigue) in M. cephalus swimming at the minimal cost of transport (i.e. optimal swimming speed; U _opt). At each temperature, a relationship was established between swimming speed and oxygen consumption (MO₂). This allowed estimation of U _opt at 45 cm s⁻¹ (~1.12 Body Length s⁻¹). Independent of temperature, stamina at U _opt was significantly reduced in severe hypoxia, i.e. at 15% of air saturation (AS). In these conditions, oxygen supply appears therefore to be insufficient to maintain swimming, even at the low speed tested here. After the stamina test, MO₂ measured in fish tested at 15% AS was significantly higher than that measured after the test in normoxia. Therefore, we suggest that in hypoxia, fish used anaerobic metabolism to supplement swimming at U _opt, leading to an oxygen debt. Since flathead grey mullet is a hypoxia-tolerant species, it is possible that hypoxic conditions less severe than those tested here may reduce stamina at low speed in less tolerant species. In addition, we suggest that testing stamina at these speeds may be relevant in order to understand the effect of hypoxia on behavioural activities carried out at low speed, such as food searching.     

Behavioural toxicity in zebrafish (Danio rerio) exposed to waterborne zinc and chromium(VI)

This study is to investigate the behavioural responses of zebrafish to the toxic effects of waterborne zinc and chromium(VI). Swimming performance of fish was tracked by a real-time toxicity bio-monitoring system and transformed into data to calculate individual and fish school behavioural endpoints. Behavioural responses under metals were in accordance with the Stepwise Behavioural Response model. Significant and concentration-dependent effects on swimming speed and turning times were found at concentrations equal to or higher than 3.5 mg L^-1 for zinc and 0.7 mg L^-1 for chromium. Swimming height significantly declined in concentrations equal to or higher than 3.5 mg L^-1 under zinc exposure, while for the chromium treatment, it significantly increased in lower concentration exposures and decreased in higher concentration exposures. Average distance and dispersion of fish school decreased at the beginning but there were no obvious changes during the whole exposure period. Behavioural responses under metal exposures were time-dependent: avoidance (increased speed and turning times and decreased average distance and dispersion) might be the first response and followed by height changes. Changes in behavioural endpoints ‘swimming speed and height’ can be used as early stress responses for acute aquatic metal contamination as part of ecological risk assessment.     

Depth-related enzymic activities in muscle,brain and heart of deep-living pelagic marine teleosts

The activities [units^-1 wet weight tissue] of lactate dehydrogenase (LDH), pyruvate kinase (PK), malate dehydrogenase (MDH) and isocitrate dehydrogenase (IDH) in white skeletal muscle, brain and heart of 24 pelagic teleost fishes were determined. In addition, for several of the foregoing species, citrate synthase (CS) activities were examined in white skeletal muscle. In muscle, the activities of all these enzymes decrease exponentially with increasing minimum depth of occurrence of the species; this decrease closely parallels the decrease in respiratory rate found previously for these same species. The decline in enzymic activity with increasing minimum depth of occurrence is much greater than the decline in body protein content of the whole fishes, suggesting a disproportionately rapid fall in enzyme concentration in comparison to contractile and structural protein concentrations. The similar reductions in activities of both glycolytic (LDH and PK) and citric acid cycle (CS, MDH and IDH) enzymes with depth indicate that both standard and active metabolisms of deeper-dwelling species are reduced relative to shallower-dwelling forms. There is no suggestion of increased anaerobic capacity with depth or in relation to species, occurrence in the oxygen minimum layer. In brain and heart, there is no significant decrease in enzymic activity with increasing minimum depth of occurrence. These two tissues have similar capacities for metabolism in most fishes, when comparisons are based on enzymic activity per gram wet weight of tissue.     

Variation in protein patterns within and among polychaete sibling species of the genus<Emphasis Type="Italic"> Marenzelleria</Emphasis> (Spionidae): a preliminary survey

Differences in protein patterns of the soluble protein fraction among the sibling species Marenzelleria viridis (formerly type I) and M. neglecta (formerly type II) were investigated under common environmental conditions using two-dimensional gel electrophoresis (2D-PAGE). Protein expression was determined using general protein staining with Coomassie-blue and compared with radioactive labeling of proteins. In the well-resolved region of stained gels an average of 319 protein spots for M. viridis and 241 spots for M. neglecta could be detected. High sensitivity of radiolabeling allowed separation of an average of 517 and 496 spots for M. viridis and for M. neglecta, respectively. Differences in protein expression between both species could be attributed mainly to qualitative differences in protein patterns. Triplet pattern was used to calculate the genetic similarity of the two species. Thus, 373 protein spots were scored for this analysis; whereas 304 spots were invariant, 36 spots were specific for M. viridis, while 33 spots were specific for M. neglecta. The genetic similarity (F) of the two Marenzelleria sibling species was 0.815. Apart from presence and absence, differences between both species resulted either from slight changes in the isoelectric point or from molecular weight, but rarely from both. Genetic variability was found only among specimens of M. viridis. The experimental conditions to perform two-dimensional electrophoresis for these polychaete species were established for subsequent investigations on a proteomic level. Using 2D-PAGE we expect further insight into the evolutionary adaptation in Marenzelleria spp.Communicated by O. Kinne, Oldendorf/Luhe