Hans Erik Karlsen

Low frequency hearing in cephalopods

SummaryClassical conditioning was employed to test the sensitivity of cephalopods to vibrations between 1 and 100 Hz generated in a standing wave acoustic tube. The animals were trained to associate sound stimuli with a weak electric shock, and the recorded conditioned responses were changes in breathing and jetting activity. Five specimens of Sepia officinalis were tested, and all responded to these low frequency sounds. The relevant stimulus parameter was particle motion rather than sound pressure. The threshold values (measured as particle acceleration) decreased towards lower frequencies in the tested range, reaching values below 4 × 10-3 m/s2. The thresholds in the most sensitive range may have been masked by the considerable background noise at the experimental site (Naples). Two individuals of Octopus vulgaris and one Loligo vulgaris were also tested, and showed a similar sensitivity to low frequency sound.

Avoidance responses to infrasound in downstream migrating European silver eels, (Anguilla anguilla)

In an attempt to develop an efficient acoustic fish fence, we have designed an infrasound source able to generate large nearfield particle acceleration. The source generates water movements by means of two symmetrical pistons in an air-filled cylinder with 21 cm bore. The pistons are driven by eccentric coupling to an electric motor, with 5 cm p.p. amplitude. The piston movements are 180° out of phase. The piston reaction forces are thus opposed, leading to vibration free operation. The submergible infrasound source is operated freely suspended in the water mass. The emitted sound frequency is 11.8 Hz. The particle acceleration is about 0.01 m s−2 at a distance of 3 m, corresponding to the threshold intensity for deterring effects of infrasound on Atlantic salmon smolts. The sound source was employed to test the effect of intense infrasound on migrating European silver eels. Fish confined in a tank displayed startle behaviour and prolonged stress reactions, telemetrically monitored as tachycardia, in response to intense infrasound. The field tests were carried out in the River Imsa. A trap that catches all the descending eels is installed near the river mouth. The trap was separated in four equal sections. During the periods with infrasound exposure, the proportion of silver eels entering the section closest to the sound source was reduced to 43% of the control value. In the section closest to the opposite river bank, infrasound increased the proportion of trapped eels to 144% of the control values. This shift of the migrating eels away from the infrasound source was highly significant.

Validation of reference genes for quantitative RT-qPCR studies of gene expression in Atlantic cod (Gadus morhua l.) during temperature stress.

BackgroundOne important physiological response to environmental stress in animals is change in gene expression. To obtain reliable data from gene expression studies using RT-qPCR it is important to evaluate a set of possible reference genes as normalizers for expression. The expression of these candidate genes should be analyzed in the relevant tissues during normal and stressed situations. To find suitable reference genes it was crucial that the genes were stably expressed also during a situation of physiological stress. For poikilotermic animals like cod, changes in temperature are normal, but if the changes are faster than physiological compensation, the animals respond with typical stress responses. It has previously been shown that Atlantic cod show stress responses when elevation of water temperature is faster than 1 degree/day, for this reason we chose hyperthermia as stress agent for this experiment.FindingsWe here describe the expression of eight candidate reference genes from Atlantic cod (Gadus morhua l.) and their stability during thermal stress (temperature elevation of one degree C/day for 5 days). The genes investigated were: Eukaryotic elongation factor 1 alpha, ef1a; 18s ribosomal RNA; 18s, Ubiquitin conjugate protein; ubiq, cytoskeletal beta-actin; actb, major histcompatibility complex I; MHC-I light chain, beta-2 -microglobulin; b2m, cytoskeletal alpha-tubulin; tba1c, acidic ribosomal phosphoprotein; rplp1, glucose-6-phosphate dehydrogenase; g6pd. Their expression were analyzed in 6 tissues (liver, head kidney, intestine, spleen, heart and gills) from cods exposed to elevated temperature and compared to a control group. Although there were variations between tissues with respect to reference gene stability, four transcripts were more consistent than the others: ubiq, ef1a, 18s and rplp1. We therefore used these to analyze the expression of stress related genes (heat shock proteins) induced during hyperthermia. We found that both transcripts were significantly upregulated in several tissues in fish exposed to increased temperature.ConclusionThis is the first study comparing reference genes for RT-qPCR analyses of expression during hyperthermia in Atlantic cod. ef1a, 18s, rplp1 and ubiq transcripts were found to be well suited as reference genes during these experimental conditions.

Infrasound initiates directional fast-start escape responses in juvenile roach Rutilus rutilus.

SUMMARY Acoustic stimuli within the sonic range are effective triggers of C-type escape behaviours in fish. We have previously shown that fish have an acute sensitivity to infrasound also, with acceleration thresholds in the range of 10–5 m s–2. In addition, infrasound at high intensities around 10–2 m s–2 elicits strong and sustained avoidance responses in several fish species. In the present study, the possible triggering of C-escapes by infrasonic single-cycle vibrations was examined in juvenile roach Rutilus rutilus. The fish were accelerated in a controlled and quantifiable manner using a swing system. The applied stimuli simulated essential components of the accelerations that a small fish would encounter in the hydrodynamic flow field produced by a predatory fish. Typical C- and S-type escape responses were induced by accelerations within the infrasonic range with a threshold of 0.023 m s–2 for an initial acceleration at 6.7 Hz. Response trajectories were on average in the same direction as the initial acceleration. Unexpectedly, startle behaviours mainly occurred in the trailing half of the test chamber, in which the fish were subjected to linear acceleration in combination with compression, i.e. the expected stimuli produced by an approaching predator. Very few responses were observed in the leading half of the test chamber, where the fish were subjected to acceleration and rarefaction, i.e. the stimuli expected from a suction type of predator. We conclude that particle acceleration is essential for the directionality of the startle response to infrasound, and that the response is triggered by the synergistic effects of acceleration and compression.

Modeling Propagation of Seismic Airgun Sounds and the Effects on Fish Behavior

High activity of seismic surveying in Norwegian waters has caused concerns about the impact the acoustic noise from the seismic airguns may have on marine life. There is evidence that this noise can cause reactions on the behavior of the fish resulting in reduced catches. To mitigate the problem and the conflict of interest between the fishing industry and the seismic exploration interest, the Norwegian Petroleum Directorate (NPD) commissioned SINTEF Information and Communication Technology (ICT, Trondheim, Norway) and the Department of Biology, University of Oslo (Oslo, Norway) to develop an acoustic-biological model to predict the impact of seismic noise on the fish population. The ultimate goal is to develop an acoustic-biological model to use in the design and planning of seismic surveys such that the disturbance to fishing interest is minimized. This acoustic module of the model is based on ray theory and can deal with range-dependent bathymetry and depth-dependent sound-speed profiles. The bottom is modeled as a sedimentary fluid layer over a solid elastic rock and the model requires the thickness and seismoacoustic properties of the sediments layer and the rock with compressional speed, shear speed, and absorption. The model simulates the total sound field, both in the time domain and in the frequency domain, out to very large distances. Calculated sound exposure levels are compared with startle response levels for cod. Preliminary conclusions indicate a required distance in the range of 5-10 km, but dependent on the depth and the season. In additions, under certain conditions, there will appear regions with hot spots where the sound level is significantly higher due to caustics and focusing of sound. Modeled results are compared with results obtained from a joint seismoacoustic survey conducted in summer 2009 at Vesterålen-Lofoten area (Nordland VII). In this experiment, signals were recorded at fixed hydrophone positions as the seismic vessel approached from a maximum distance of 30 km toward the receiving positions. The same situation was modeled using available geological and oceanographic information as input to the acoustic model. The agreement between the real and recorded signals and the model results is good. This indicates that in the future acoustic-biological models may be used in the design and planning of seismic surveys such that the disturbance to fishing is minimized.

Effect of Diamphidia toxin, a bushman arrow poison, on ionic permeability in nucleated cells

The effect of Diamphidia toxin, isolated from pupae of Diamphidia nigro-ornata, was tested on two different cell lines (GH4C1 cells and HL-60 cells) and on human lymphocytes. The toxin raised intracellular Ca2+ concentration, as assessed with quin 2, in a dose-related manner in all three cell types. The effect was abolished when extracellular Ca2+ was chelated by EGTA. Low concentrations of the toxin evoked a delayed as well as a smaller response. The response time was also temperature-dependent, with a Q10 of about 2. Low, but effective concentrations of the toxin did not affect cell membrane integrity, as tested with Trypan blue, and induced a seemingly physiological release of prolactin from the GH4C1 cells. Diamphidia toxins effect on the membrane permeability of GH4C1 cells was further investigated with patch-clamp techniques. The toxin appeared to increase the conductance for all small ions without affecting the normal ionic channels present in these cells. We conclude that Diamphidia toxin has a general effect on the plasma membrane of different cell types and thereby increases, probably non-specifically, the permeability for small ions.

Behavioural responses to infrasonic particle acceleration in cuttlefish

ABSTRACT Attacks by aquatic predators generate frontal water disturbances characterised by low-frequency gradients in pressure and particle motion. Low-frequency hearing is highly developed in cephalopods. Thus, we examined behavioural responses in juvenile cuttlefish to infrasonic accelerations mimicking main aspects of the hydrodynamic signals created by predators. In the experimental set-up, animals and their surrounding water moved as a unit to minimise lateral line activation and to allow examination of the contribution by the inner ear. Behavioural responses were tested in light versus darkness and after food deprivation following a ‘simulated’ hunting opportunity. At low acceleration levels, colour change threshold at 3, 5 and 9 Hz was 0.028, 0.038 and 0.035 m s−2, respectively. At higher stimulus levels, jet-propulsed escape responses thresholds in daylight were 0.043, 0.065 and 0.069 m s−2 at 3, 5 and 9 Hz, respectively, and not significantly different from the corresponding darkness thresholds of 0.043, 0.071 and 0.064 m s−2. In a simulated hunting mode, escape thresholds were significantly higher at 3 Hz (0.118 m s−2) but not at 9 Hz (0.134 m s−2). Escape responses were directional, and overall followed the direction of the initial particle acceleration, with mean escape angles from 313 to 33 deg for all three experiments. Thus, in the wild, particle acceleration might cause escape responses directed away from striking predators but towards suction-feeding predators. We suggest that cuttlefish jet-propulsed escape behaviour has evolved to be elicited by the early hydrodynamic disturbances generated during predator encounters, and that the inner ear plays an essential role in the acoustic escape responses. Editors’ Choice: Cuttlefish exhibit jet-propulsed escape responses adapted to the hydrodynamic signatures generated by predators in the initial approach phase of an attack.