Computational methods for the analysis of swimming biomechanics

A series of computer programs is presented which enables the analysis of fish body shape and mass distributions, spine positions, spine curvatures and coordinates for the centre of mass. Data are derived from silhouette outlines of swimming fish, white muscle strains during swimming, white muscle force-time development functions for body bending cycles, muscle force and power production along the whole fish body and hydrodynamic efficiencies for fast-start swimming behaviours.     

Circannual pupation rhythm in the varied carpet beetle Anthrenus verbasci under different nutrient conditions

Anthrenus verbasci pupates in spring and the timing of pupation is controlled by a circannual rhythm. Although A. verbasci is considered to be a univoltine species in Japan, it is assumed that larval development in its natural habitats, including bird nests, varies with nutrient availability, and that the life cycle often takes two or more years to complete. In the present study, larval development and pupation times were compared under constant and outdoor conditions in larvae provided a diet of either high‐nutrient bonito powder or low‐nutrient pigeon feathers. Although a circannual pupation rhythm was observed irrespective of the diet used, larval development was slower on feathers than on bonito powder. The pupation times on feathers varied over three years or more under both constant and outdoor conditions. Under outdoor conditions, larvae grown on feathers needed three years to approach the weight gained within a year by larvae grown on bonito powder. It is considered that life cycle length in A. verbasci is often two years or more in nutritionally unstable natural habitats, and that this species has probably evolved a circannual rhythm as a seasonal adaptation to nutrient‐poor environments.     

Effect of the bonito insulin on the insulin release from monolayer culture of rat pancreas.

The effect of bonito insulin on insulin release was examined in the monolayer culture of rat pancreatic beta-cells. The beta-cells were preincubated for 5 to 20 hr with or without a small dose (100 microunits/ml) of bonito insulin in the medium containing 100 mg% glucose. And then, they were incubated in 300 mg% glucose alone or together with bonito insulin for 5 hr. There was no significant difference between the IRI release from these beta-cells with or without bonito insulin. The concentration of bonito insulin was augmented from 100 microunits/ml to 500, 1,000 and 2,000 microunits/ml. A significant inhibitory effect on the glucose-induced insulin release was observed only after the preincubation for 20 hr with 2,000 microunits/ml of bonito insulin.     

The effect of substratum type on aspects of swimming performance and behaviour in shortnose sturgeon Acipenser brevirostrum

The swimming performance and associated swimming behaviour (i.e. substratum‐skimming, station‐holding and free swimming) were assessed in shortnose sturgeon Acipenser brevirostrum during critical swimming and endurance swimming tests over a rough and a smooth substratum. It was hypothesized that the addition of a rough substratum in the swimming flume may provide a surface for the A. brevirostrum to grip and offer an energetic advantage. Substratum type did not affect the critical swimming performance, but A. brevirostrum consistently performed more bottom behaviours (i.e. substratum‐skimming and station‐holding) while on a smooth substratum. Acipenser brevirostrum had little contact with the rough substratum until the velocity was >1 body length s^?1. Endurance swimming time was significantly lower for A. brevirostrum over the rough bottom at the highest velocity (30 cm s^?1) which may be attributed to the observed increase in free swimming and decrease in bottom behaviours. During endurance swimming, the rough substratum was mainly used at intermediate velocities, suggesting that there may be a stability cost associated with being in contact with the rough substratum at certain velocities.     

Copepod flow modes and modulation: a modelling study of the water currents produced by an unsteadily swimming copepod

Video observation has shown that feeding-current-producing calanoid copepods modulate their feeding currents by displaying a sequence of different swimming behaviours during a time period of up to tens of seconds. In order to understand the feeding-current modulation process, we numerically modelled the steady feeding currents for different modes of observed copepod motion behaviours (i.e. free sinking, partial sinking, hovering, vertical swimming upward and horizontal swimming backward or forward). Based on observational data, we also reproduced numerically a modulated feeding current associated with an unsteadily swimming copepod. We found that: (i) by changing its propulsive force, a copepod can switch between different swimming behaviours, leading to completely different flow-field patterns in self-generated surrounding flow; (ii) by exerting a time-varying propulsive force, a copepod can modulate temporally the basic flow modes to create an unsteady feeding current which manipulates precisely the trajectories of entrained food particles over a long time period; (iii) the modulation process may be energetically more efficient than exerting a constant propulsive force onto water to create a constant feeding current of a wider entrainment range. A probable reason is that the modulated unsteady flow entrains those water parcels containing food particles and leaves behind those without valuable food in them.     

Structure of bonito heart ferricytochrome c and some remarks on molecular interaction in its crystalline state.

The structure analysis of bonito heart ferricytochrome c was carried out at 2.8 A resolution by X-ray diffraction. The overall features of the molecule are virtually identical with those of bonito ferrocytochromes c and other cytochromes c. In the present work, the modes of molecular packing among cytochromes c were also compared by means of intermolecular distance maps. Some differences in the structures of ferro- and ferricytochrome c may exist on the surface of the molecules.     

Isolation and characterization of growth hormone from a marine fish, bonito (Katsuwonus pelamis)

Growth hormone (GH) was extracted under alkaline conditions (pH 10) from pituitary glands (6.3 g) of bonito (Katsuwonus pelamis), and subsequently purified by gel filtration, ion exchange chromatography, and reversed-phase HPLC. The GH was monitored by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and by immunoblotting with yellowtail GH antiserum at each step of purification. GH activity was determined by an in vivo bioassay. The yield of this hormone was 4.8 mg/g wet tissue. Intraperitoneal injection of bonito GH at doses of 0.1 and 1 micrograms/g body wt at 7-day intervals resulted in a significant increase in body weight and length of juvenile rainbow trout. Bonito GH antiserum exhibited both species and hormone specificity in radioimmunoassay. However, the bonito GH antiserum as well as yellowtail GH antiserum exhibited hormone specificity but not species specificity in immunoblotting. A molecular weight of 21,000 and an isoelectric point of 7.0 for bonito GH were estimated by SDS-PAGE and gel electrofocusing, respectively. The complete amino acid sequence of 185 residues was determined by sequencing fragment peptides prepared by chemical and enzymatic cleavages. Sequence comparison of bonito GH with other GHs revealed that there is a significant deletion in the middle of the molecule.     

Nano-tags for neonates and ocean-mediated swimming behaviours linked to rapid dispersal of hatchling sea turtles

Dispersal during juvenile life stages drives the life-history evolution and dynamics of many marine vertebrate populations. However, the movements of juvenile organisms, too small to track using conventional satellite telemetry devices, remain enigmatic. For sea turtles, this led to the paradigm of the ‘lost years'' since hatchlings disperse widely with ocean currents. Recently, advances in the miniaturization of tracking technology have permitted the application of nano-tags to track cryptic organisms. Here, the novel use of acoustic nano-tags on neonate loggerhead turtle hatchlings enabled us to witness first-hand their dispersal and behaviour during their first day at sea. We tracked hatchlings distances of up to 15 km and documented their rapid transport (up to 60 m min⁻¹) with surface current flows passing their natal areas. Tracking was complemented with laboratory observations to monitor swimming behaviours over longer periods which highlighted (i) a positive correlation between swimming activity levels and body size and (ii) population-specific swimming behaviours (e.g. nocturnal inactivity) suggesting local oceanic conditions drive the evolution of innate swimming behaviours. Knowledge of the swimming behaviours of small organisms is crucial to improve the accuracy of ocean model simulations used to predict the fate of these organisms and determine resultant population-level implications into adulthood.     

Cardiorespiratory performance during prolonged swimming tests with salmonids: a perspective on temperature effects and potential analytical pitfalls

A prolonged swimming trial is the most common approach in studying steady-state changes in oxygen uptake, cardiac output and tissue oxygen extraction as a function of swimming speed in salmonids. The data generated by these sorts of studies are used here to support the idea that a maximum oxygen uptake is reached during a critical swimming speed test. Maximum oxygen uptake has a temperature optimum. Potential explanations are advanced to explain why maximum aerobic performance falls off at high temperature. The valuable information provided by critical swimming tests can be confounded by non-steady-state swimming behaviours, which typically occur with increasing frequency as salmonids approach fatigue. Two major concerns are noted. Foremost, measurements of oxygen uptake during swimming can considerably underestimate the true cost of transport near critical swimming speed, apparently in a temperature-dependent manner. Second, based on a comparison with voluntary swimming ascents in a raceway, forced swimming trials in a swim tunnel respirometer may underestimate critical swimming speed, possibly because fish in a swim tunnel respirometer are unable to sustain a ground speed.     

Eye colour and aggression in juvenile guppies, Poecilia reticulata peters (Pisces: Poeciliidae)

A series of four related experiments on guppy, Poecilia reticulata, juveniles indicates that eye colour is related to motivational state. In juveniles of this species two distinct eye colour states are easily discernible, with a transient intermediate state infrequently seen. These states are dark, light, and intermediate. In aggressive interactions involving both both dark- and light-eyed fish, dark-eyed fish won 98.6% of the encounters (N=222 encounters). Eye colours are significantly correlated with bout lengths of certain behaviours (aggressive encounters, swimming, substrate biting); total time spent in certain behaviours (swimming, aggressive encounters); and type and intensity of aggressive encounters. Eye colour appears to be related to motivational state and may serve a signal function.     

The flow field around a freely swimming copepod in steady motion. Part I: Theoretical analysis

The three-dimensional flow field around a free-swimming copepodin steady motion was studiedtheoretically. This study was basedon coupling the Navier–Stokes equations with the dynamicequations for an idealized body of a copepod. To allow analyticalsolutions to the flow field, three simplifications were made:(a) to simulate the effect of the beating movement of the cephalicappendages, a force-field was added to the Navier–Stokesequations, (b) to linearize the problem, Stokes flow was used,and (c) to simplify the morphologies of the copepods, a sphericalbody shape was assumed. Analytical solutions were derived forfive steady motions: (1) hovering, (2) sinking, (3) upwardsswimming, (4) backwards swimming and (5) forwards swimming.The results show that thegeometry of the flow field around afreely swimming copepod varies significantly with the differentswimming behaviours. When a copepod hovers in the water, orswims very slowly, it generates a wide, cone-shaped flow field.In contrast, when a copepod sinks, or swims fast, the flow geometryis not cone-shaped, but cylindrical, narrow and long. Theseresults are consistent with published observations on live copepods.It is shown that the differences in the flow geometry with thedifferent swimming behaviours are due to the relative importancebetween the two factors in generating the flow field: the copepod'sswimming motion and the requirement to counterbalance the copepod'sexcess weight. The results also highlight the importance ofconsidering freely swimming copepods as self-propelled ratherthan as towed bodies. ‘Self-propelled’ means a freelyswimming copepod must gain thrust from the surrounding waterin order to counterbalance the drag force by water and its excessweight. Regardless of swimming behaviours and velocities, thefar-field velocity field decays to that of the velocity fieldgenerated by a point force of magnitude equal to the copepod'sexcess weight in an infinite domain. On the other hand, usingthe towed body model yields a flow field with much differentfar- and near-field flow characteristics. Hence, the towed bodymodel is inherently unable to reproduce fundamental characteristicsof the flow field around a freely swimming copepod.     

Ontogeny of behaviour in larvae of marine demersal fishes

The development of behaviours that are relevant to larval dispersal of marine, demersal fishes is poorly understood. This review focuses on recent work that attempts to quantify the development of swimming, orientation, vertical distribution and sensory abilities. These behaviours are developed enough to influence dispersal outcomes during most of the pelagic larval stage. Larvae swim in the ocean at speeds similar to the currents found in many locations and at 3–15 body lengths per second (BL s⁻¹), although, based on laboratory measurements, species from cold environments swim slower than those from warm environments. At least in warm-water species, larvae swim in an inertial hydrodynamic environment for most of their pelagic period. Unfed swimming endurance is >10 km from about 8–10 mm, and reaches more than 50 km before settlement in several species. Larval fishes are efficient swimmers. In most species, a large majority of larvae have orientated swimming in the ocean, but the precision of orientation does not improve with growth. Swimming direction of the larvae frequently changes ontogenetically. Vertical distribution changes ontogenetically in most species, and both ontogenetic ascents and descents are found. Development of schooling is poorly understood, but it may influence speed, orientation and vertical distribution. Sensory abilities (hearing, olfaction, vision) form early, are well developed and are able to detect cues relevant to orientation for most of the pelagic larval stage. All this indicates that the passive portion of the pelagic larval duration will be short, at least in most warm-water species, and that behaviour must be taken into account when considering dispersal, and in particular in dispersal models. Although quantitative information on the ontogeny of some behaviours is available for a relatively small number of species, more research in this field is required, especially on species from colder waters.     

Intake of dried bonito broth flavored with dextrin solution induced conditioned place preference in mice

We investigated to determine whether dried bonito broth flavor induces a reinforcing effect using the conditioned place preference (CPP) test. Only dried bonito broth did not induce CPP. Sucrose induced CPP in 20% solution. A 21.86% dextrin solution, with the same calorie content as the 20% sucrose solution, did not induce CPP, but a dextrin solution flavored with dried bonito broth (BD) induced CPP. An AD solution containing the same concentrations of dextrin, NaCl, IMP, GMP, and amino acids as found in BD tended to increase the time spent in the conditioned box but did not significantly. Aromatic compounds, such as citral, vanillin, and menthol flavored AD solutions did not induce CPP, whereas an AD solution supplemented with dried bonito flavoring agent induced CPP. In mice with transected olfactory nerves, CPP was not induced by voluntary intake of BD. These results suggest that the aromatic profile of the dried bonito broth plays an important role in BD-induced CPP.     

Group swimming behaviour and energetics in bluegill Lepomis macrochirus and rainbow trout Oncorhynchus mykiss

Group swimming size influences metabolic energy consumption and swimming behaviour in fishes. Hydrodynamic flows and vortices of other fish are thought to be beneficial in terms of the energetic costs of swimming. Similarly, abiotic obstructions have been shown to have similar benefits with respect to metabolic consumption in swimming fish such as rainbow trout Oncorhynchus mykiss. The current study works to examine metabolic rates and swimming behaviours as a function of group swimming with bluegill sunfish Lepomis macrochirus and O. mykiss. Fishes were subjected to individual and group swimming in a respiratory swim tunnel to determine oxygen consumption as a proxy for the metabolic rate of swimming fish. In addition, fish movements within the swim tunnel test chamber were tracked to examine group swimming behaviour. We hypothesized that fish would benefit metabolically from group swimming. In the case of O. mykiss, we also hypothesized that groups would benefit from the presence of an abiotic structure, as has been previously observed in fish swimming individually. Our results suggest that the influence of group size on swimming metabolism is species specific. While L. macrochirus show decreased metabolic rate when swimming in a group compared to individually, O. mykiss did not show such a metabolic benefit from group swimming.     

The effect of predation risk on the predatory behaviour of a sit-and-wait predator,Ranatra dispar (Heteroptera: Nepidae), the water stick insect

Two primary defence behaviours, fore-leg extension to enhance crypsis and swimming to bottom and remaining motionless, of a predatory water bug,Ranatra dispar, are described along with their subsequent effect on foraging behaviour. It was hypothesised that hungry predators would respond less and for a shorter duration compared with recently fed individuals when exposed to a model threat stimulus, thereby tending to take a higher risk of predation during feeding than satiated animals. A greater proportion of animals responded overall with the leg extension response compared with the swimming response, although the mean duration of the former was significantly shorter than the latter response. A significantly higher proportion of nonfasted predators responded, and for a longer duration than fasted individuals. The type of model used significantly effected the proportion of animals that responded with leg extensions but not on its duration. In contrast, both number of animals and the duration of the swim/motionless behaviour were significantly effected by stimulus type. The subsequent effect of these 2 behavioural responses on feeding behaviour was examined and showed that although about the same number of predators removed prey from their mouthparts during both responses, significantly more prey were dropped, and therefore lost, during swimming. The results clearly indicate the significant effects that defensive behaviours have on time budgets in foraging behaviour.     

Preference for Dried Bonito Broth in Olfactory-Blocked or Taste Nerve-Sectioned Mice in the Two-Bottle Choice Test

We investigated how gustatory and olfactory information contributes to the preference for dried bonito broth in mice. In the two-bottle preference test, intact mice consumed dried bonito broth in preference to water or an amino acid-nucleotide (AN) solution containing the same concentration of amino acids and nucleotides as that in dried bonito broth. It was observed that mice with transected bilateral chorda tympani (CT) nerves, those with transected bilateral glossopharyngeal (GL) nerves, and those that were intranasally administered with zinc sulfate preferred dried bonito broth to water. Zinc sulfate was used to produce a temporary loss of olfaction. In the two-bottle preference test with dried bonito broth and an AN solution, the preference for the former was reduced in mice with transected bilateral GL nerves and in those with an olfactory blockade, but not in mice with transected bilateral CT nerves. These results suggest that dried bonito broth was preferred over the AN solution, and that simultaneous inputs from olfaction and the GL nerve contributed to this preference.     

Preference for dried bonito broth in olfactory-blocked or taste nerve-sectioned mice in the two-bottle choice test

We investigated how gustatory and olfactory information contributes to the preference for dried bonito broth in mice. In the two-bottle preference test, intact mice consumed dried bonito broth in preference to water or an amino acid-nucleotide (AN) solution containing the same concentration of amino acids and nucleotides as that in dried bonito broth. It was observed that mice with transected bilateral chorda tympani (CT) nerves, those with transected bilateral glossopharyngeal (GL) nerves, and those that were intranasally administered with zinc sulfate preferred dried bonito broth to water. Zinc sulfate was used to produce a temporary loss of olfaction. In the two-bottle preference test with dried bonito broth and an AN solution, the preference for the former was reduced in mice with transected bilateral GL nerves and in those with an olfactory blockade, but not in mice with transected bilateral CT nerves. These results suggest that dried bonito broth was preferred over the AN solution, and that simultaneous inputs from olfaction and the GL nerve contributed to this preference.     

Collective behaviours in organoids

Collective behaviour emerges from interacting units within communities, such as migrating herds, swimming fish schools, and cells within tissues. At the microscopic level, collective behaviours include collective cell migration in development and cancer invasion, rhythmic gene expression in pattern formation, cell competition in homeostasis and cancer, force generation and mechano-sensing in morphogenesis. Studying the initiation and the maintenance of collective cell behaviours is key to understand the principles of development, regeneration and disease. However, the manifold influences of contributing factors in in vivo environments challenge the dissection of causalities in animal models. As an alternative model that has emerged to overcome this difficulty, in vitro three-dimensional organoid cultures provide a reductionist approach yet retain similarities with the in vivo tissue in cellular composition and tissue organisation. Here, we focus on recent progresses in studying collective behaviours in different organoid systems and discuss their advantages and the possibility of improvement for future applications.     

Mutagenicity of smoked, dried bonito products

Mutagens have been found in smoked, dried bonito products, popular items in Japanese foods. The mutagens were isolated by means of blue cotton, an absorbent cotton preparation with covalently bound trisulfo-copper-phthalocyanine residues, and by means of XAD-2 resin. The mutagenicity was positive in Salmonella typhimurium strain TA98 with metabolic activation. The mutagens are produced during the process of smoking-and-drying bonito (a process called baikan). The activity was much higher than that expected from the content of benzo[a]pyrene. In contrast to benzo[a]pyrene, the mutagens were not inhibited by ellagic acid. The mutagenicity was not abolished by treatment with nitrite. Thin-layer and high-performance liquid chromatographic analysis gave two mutagenic fractions, both of which were distinguishable from benzo[a]pyrene and from the pyrolysis products Trp-P-1, Trp-P-2, Glu-P-1, Glu-P-2, A alpha C and MeA alpha C. The major mutagenic component was not chromatographically distinguishable from IQ and MeIQx, and the minor one was very similar to MeIQ. The smoked, dried bonito products contained free fatty acids, which were inhibitory to the mutagenicity of the bonito products.     

Effects of food type on diel behaviours of common carp Cyprinus carpio in simulated aquaculture pond conditions

In order to better understand behaviour patterns of common carp Cyprinus carpio in aquaculture ponds, their diel grazing, swimming, resting and schooling behaviours were observed in six 1 m² tanks under simulated pond conditions. Each tank was fertilized to stimulate natural food production before starting experiments, and then stocked with three C. carpio . Fish behaviours were compared among three treatments: (1) tanks with plankton only, (2) tanks with plankton and benthic macroinvertebrates and (3) tanks with plankton, benthic macroinvertebrates and artificial feed. Overall C. carpio grazed more frequently during daytime than at night and exhibited the reverse pattern for non-feeding swimming behaviour. A significant negative relationship ( r ² = 0·99, P < 0·01, n = 48) was observed between total per cent grazing time and total per cent swimming time. Fish dispersed to graze individually during daytime but schooled at night and did not display any agonistic behaviours. Diel variations in the vertical swimming behaviour of C. carpio were related to food types available. In tanks containing plankton only, fish grazed in the water column, whereas when benthic macroinvertebrates were present, they spent more time near the tank bottom. Resting behaviour was only seen in tanks with artificial feed and even then was rare (2–5% of total time). Results suggest that C. carpio growth and feed utilization efficiency in semi-intensive aquaculture systems could be optimized by using a combination of plankton, benthic macroinvertebrates and artificial feed, and feeding fish twice per day (at c . 0730 and c . 1630 hours).