Spontaneous modulations of the electric organ discharge in the weakly electric fish, Apteronotus leptorhynchus: a biophysical and behavioral analysis

Brown ghosts, Apteronotus leptorhynchus, are weakly electric gymnotiform fish whose wave-like electric organ discharges are distinguished by their enormous degree of regularity. Despite this constancy, two major types of transient electric organ discharge modulations occur: gradual frequency rises, which are characterized by a relatively fast increase in electric organ discharge frequency and a slow return to baseline frequency; and chirps, brief and complex frequency and amplitude modulations. Although in spontaneously generated gradual frequency rises both duration and amount of the frequency increase are highly variable, no distinct subtypes appear to exist. This contrasts with spontaneously generated chirps which could be divided into four "natural" subtypes based on duration, amount of frequency increase and amplitude reduction, and time-course of the frequency change. Under non-evoked conditions, gradual frequency rises and chirps occur rather rarely. External stimulation with an electrical sine wave mimicking the electric field of a neighboring fish leads to a dramatic increase in the rate of chirping not only during the 30 s of stimulation, but also in the period immediately following the stimulation. The rate of occurrence of gradual frequency rises is, however, unaffected by such a stimulation regime.     

From oscillators to modulators: behavioral and neural control of modulations of the electric organ discharge in the gymnotiform fish, Apteronotus leptorhynchus.

The brown ghost (Apteronotus leptorhynchus) is a weakly electric gymnotiform fish that produces wave-like electric organ discharges distinguished by their enormous degree of regularity. Transient modulations of these discharges occur both spontaneously and when stimulating the fish with external electric signals that mimic encounters with a neighboring fish. Two prominent forms of modulations are chirps and gradual frequency rises. Chirps are complex frequency and amplitude modulations lasting between 20 ms and more than 200 ms. Based on their biophysical characteristics, they can be divided into four distinct categories. Gradual frequency rises consist of a rise in discharge frequency, followed by a slow return to baseline frequency. Although the modulatory phase may vary considerably between a few 100 ms and almost 100 s, there is no evidence for the existence of distinct categories of this type of modulation signal. Stimulation of the fish with external electric signals results almost exclusively in the generation of type-2 chirps. This effect is independent of the chirp type generated by the respective individual under non-evoked conditions. By contrast, no proper stimulation condition is known to evoke the other three types of chirps or gradual frequency rises in non-breeding fish. In contrast to the type-2 chirps evoked when subjecting the fish to external electric stimulation, the rate of spontaneously produced chirps is quite low. However, their rate appears to be optimized according to the probability of encountering a conspecific. As a result, the rate of non-evoked chirping is increased during the night when the fish exhibit high locomotor activity and in the time period following external electric stimulation. These, as well as other, observations demonstrate that both the type and rate of modulatory behavior are affected by a variety of behavioral conditions. This diversity at the behavioral level correlates with, and is likely to be causally linked to, the diversity of inputs received by the neurons that control chirps and gradual frequency rises, respectively. These neurons form two distinct sub-nuclei within the central posterior/prepacemaker nucleus in the dorsal thalamus. In vitro tract-tracing experiments have elucidated some of the connections of this complex with other brain regions. Direct input is received from the optic tectum. Indirect input arising from telencephalic and hypothalamic regions, as well as from the preoptic area, is relayed to the central posterior/prepacemaker nucleus via the preglomerular nucleus. Feedback loops may be provided by projections of the central posterior/prepacemaker nucleus to the preglomerular nucleus and the nucleus preopticus periventricularis.     

Electrocommunication signals in female brown ghost electric knifefish, Apteronotus leptorhynchus

Female communication behaviors are often overlooked by researchers in favor of male behaviors, which are usually more overt and easier to elicit. Very little is known about female electrocommunication behaviors in brown ghost knifefish, a weakly electric wavetype Gymnotiform fish. Most behavioral studies have focused on males, and fish are usually restrained and played a stimulus near their own electric organ discharge frequency to evoke chirps (abrupt short-term frequency rises) or the jamming avoidance response. Our study focuses on categorizing and describing spontaneous and evoked electric organ discharge modulations in free-swimming female fish that were either electrically coupled to tanks containing a conspecific (male or female), or left isolated. Cluster analysis of signals produced under isolated and social conditions revealed three categories of rises: short rise, medium rise and long rise; and one category of frequency decrease (dip). Females produce significantly more short rises when electrically coupled to tanks containing lower-frequency females, and produce more long rises when electrically coupled to tanks containing males. Short rises may have an intrasexual aggressive function, while long rises may serve as an advertisement of status or reproductive condition in intersexual interactions.     

From distributed sensory processing to discrete motor representations in the diencephalon of the electric fish,Eigenmannia

Summary During their jamming avoidance response (JAR), weakly electric fish of the genusEigenmannia shift their electric organ discharge (EOD) frequency away from a similar EOD frequency of a neighboring fish. The behavioral rules and neural substrates for stimulus recognition and motor control of the JAR have been extensively studied (see review by Heiligenberg 1986). The diencephalic nucleus electrosensorius (nE) links sensory processing within the torus semicircularis and optic tectum with the mesencephalic prepacemaker nucleus which, in turn, modulates the medullary pacemaker nucleus and hence the EOD frequency. Two separate areas within the nE responsible for JAR-related EOD frequency rises and frequency falls, respectively, were identified by iontophoresis of the excitatory amino acid L-glutamate. Bilateral lesion of the areas causing EOD frequency rises resulted in elimination of JAR-related frequency rises above a baseline frequency obtained in the absence of a jamming stimulus. Similarly, bilateral lesion of the areas causing frequency falls resulted in a loss of JAR-related frequency falls below the baseline frequency. Whether these areas are also responsible for non-JAR-related frequency shifts is not known. The strength of response and spatial extent of the areas causing frequency shifts varied among fish and also varied in individual fish, reflecting the strength of JAR-related frequency shifts and the balance of activities in frequency-rise and frequency-fall areas. Local application of bicuculline-methiodide or GABA demonstrated a tonic inhibitory input to each area and suggests a reciprocal inhibitory interaction between the two ipsilateral areas, possibly accounting for much of the individual plasticity.The nE thus is a site for neuronal transformation from distributed, topographically organized processing within the laminated structures of the torus and tectum to discrete cell clusters which control antagonistic motor responses.Abbreviations EOD electric organ discharge - JAR jamming avoidance response - Df difference frequency between jamming signal and the fish's own EOD - nE nucleus electrosensorius - PPn prepacemaker nucleus     

Sensory cues for the gradual frequency fall responses of the gymnotiform electric fish, Rhamphichthys rostratus

The sensory cues for a less known form of frequency shifting behavior, gradual frequency falls, of electric organ discharges (EODs) in a pulse-type gymnotiform electric fish, Rhamphichthys rostratus, were identified. We found that the gradual frequency fall occurs independently of more commonly observed momentary phase shifting behavior, and is due to perturbation of sensory feedback of the fish's own EODs by EODs of neighboring fish. The following components were identified as essential features in the signal mixture of the fish's own and the neighbor's EOD pulses: (1) the neighbor's pulses must be placed within a few millisecond of the fish's own pulses, (2) the neighbor's pulses, presented singly at low frequencies (0.2–4 Hz), were sufficient, (3) the frequency of individual pulse presentation must be below 4 Hz, (4) amplitude modulation of the sensory feedback of the fish's own pulses induced by such insertions of the neighbor's pulses must contain a high frequency component: sinusoidal amplitude modulation of the fish's own EOD feedback at these low frequencies does not induce gradual frequency falls. Differential stimulation across body surfaces, which is required for the jamming avoidance response (JAR) of wave-type gymnotiform electric fish, was not necessary for this behavior. We propose a cascade of high-pass and low-pass frequency filters within the amplitude processing pathway in the central nervous system as the mechanism of the gradual frequency fall response.Abbreviations EOD electric organ discharge - f frequency of EOD or pacemaker command signal - JAR jamming avoidance response - S ₁ stimulus mimicking fish's own EOD - f ₁ frequency of S₁ - S ₂ stimulus mimicking neighbor's EOD - f ₂ frequency of S₂     

The consequences of a drastic fish stock reduction in the large and shallow Lake Wolderwijd,The Netherlands. Can we understand what happened?

In 1990 an experiment started in the large and shallow lake Wolderwijd (2700 ha, mean depth 1.5 m) to improve the water quality. About 75% of the fish stock was removed (425 000 kg fish). The fish was mainly composed of bream and roach. In May 600000 young pikes (3–4 cm) were introduced.In May 1991 the water became very clear (Secchi depth 1.8 m) during a spring bloom of large Daphnia. Then the grazing by zooplankton was eight times higher than the primary production of algae and the total suspended matter concentration became very low. Compared to the situation before the fish reduction, the grazing had increased only slightly, while the primary production had decreased significantly in early spring. The fish stock reduction might have contributed to the reduction in primary production by a reduced internal nutrient load. The clear water period lasted six weeks. Daphnia disappeared in July due to food limitation, the algal biomass increased and the Secchi depth became 50 cm. Daphnia did not recover during summer, due to predation that was not caused by 0 + fish but by the mysid shrimp Neomysis integer. Neomysis could develop abundantly, because of the reduced biomass of the predator perch. The production of young fish had been low because of the cold spring weather. The cold weather was probably also responsible for the slow increase in density of macrophytes. After 1991, perch probably can control Neomysis. Due to lack of spawning places and shelter for 0 + pike, pike was probably not able to control the production of 0 + fish. In a lake of this scale, it will not be easy to get more than 50% coverage of macrophytes, which seems necessary to keep the algal biomass low by nutrient competition. Therefore, we expect also in the future a decrease in transparency in the summer. Locally, especially near Characeae, the water might stay clear.     

The ultrastructure of chloride cells in the gills of the teleostOreochromis mossambicus during exposure to acidified water

Summary Branchial chloride cells, which actively take up ions in the gills of freshwater fish, were studied in tilapia (Oreochromis mossambicus) exposed to sublethally acidified freshwater. Structural damage of cells, resulting in cell death by necrosis, only occurred transiently, when the reduction of water pH was acute rather than gradual. The most prominent effects of water acidification were the rapid increase in the number of chloride cells and the changes in frequency of the different stages of the chloride cell cycle. In the opercular inner epithelium, a twofold increase in cells occurred 48 h after gradual acidification. Cell density stabilized after 4 weeks at a level 5 times that of control fish. Four transitory stages were distinguished in the chloride cell cycle: accessory or replacement cells, immature, mature, and degenerating (apoptotic) cells. In control fish, mature chloride cells dominated (over 50%) with immature and apoptotic cells totalling about 40%. After 4 weeks in acid water, only 13% of the cells were mature. Immature and apoptotic cells dominated, each representing about 40% of the total number of chloride cells. Mature cells apparently age rapidly under these conditions. Thus, chloride cells turn over quickly in acid water, with a minor increase in ion transport capacity of the gills. This conclusion is supported by the observation that opercular and branchial Na⁺/K⁺ ATPase activities in treated fish are only 40%–50% higher than in controls.     

The development of the Jamming Avoidance Response (JAR) in Eigenmannia: An innate behavior indeed

Summary In its Jamming Avoidance Response (JAR), the gymnotiform electric fish Eigenmannia shifts its electric organ discharge (EOD) frequency away from similar interfering frequencies. Continual behavioral measurements were carried out in 164 juvenile fish until a correct JAR emerged. Sixty-four of these fish were raised in complete isolation, the remainder in a community of their siblings. A correct JAR emerged in fish of 1.2–1.6 cm in body length, corresponding to a developmental age of 24–32 days. In 6 of 164 fish, the emergence of a correct JAR followed an interim appearance of an incorrect JAR, which involved frequency shifts in the direction opposite to those of a correct JAR. The fish raised in isolation developed the same forms of behavior and showed the same sequence in their appearance as did socially raised fish. This indicates that the JAR and its developmental schedule are innate. The appearance of an incorrect JAR suggests initial errors or incompleteness in the wiring of central nervous connections. A correct JAR ultimately emerged even if a stimulus regimen was offered that rewarded frequency shifts in the direction opposite to those of a correct JAR. This indicates that the development of the JAR is immune to experimental alterations of sensory experience.Abbreviations Df frequency difference between a jamming stimulus and fish's EOD - ELL electrosensory lateral line lobe - EO electric organ - EOD electric organ discharge - JAR Jamming Avoidance Response - nE nucleus electrosensorius - nE subnucleus of nE, causing drop of EOD frequency - nE subnucleus of nE, causing rise of EOD frequency - Pn pacemaker nucleus - PPn prepacemaker nucleus     

Abundance and daytime vertical distribution of planktonic fish larvae in an oligotrophic South Island lake

The vertical distribution of common bully (Gobiomorphus cotidianus) and koaro (Galaxias brevipinnis) larvae in the limnetic zone of Lake Coleridge were determined using a high-frequency (200 KHz) echosounder. Planktonic bully larvae first appeared in appreciable numbers in January. By February, they formed a scattering layer between depths of 12 to 24 m during the day, where they achieved a maximum density of 0.59 fish m^-3. Larger (> 18mm) fish migrated to the littoral zone and densities declined to < 0.01 fish m^-3 by July, when remaining larval fish occupied greater daytime depths. Their vertical distribution during the day appeared to be influenced mainly by light levels and water temperatures. Larvae grew more slowly (0.12 mm d^-1) than in more productive North Island lakes, and were also present in lower densities for a more restricted period of time. Koaro larvae first appeared in November and December and were found in low numbers (< 0.01 fish m^-3) in summer at depths of 10 to 26 m. Salmonid production in the limnetic zone is probably limited by the small size and relative scarcity of forage fish present.     

Formation of morphological defences in response to YOY perch and invertebrate predation in two <Emphasis Type="Italic">Daphnia</Emphasis> species coexisting in a mesotrophic lake

This study examined the formation of morphological defences by two coexisting Daphnia species, the large-sized D. pulicaria (2 mm) and the small-sized D. mendotae (1.4 mm), in response to the presence of young-of-the-year (YOY) yellow perch (Perca flavescens) and invertebrate predators (Chaoborus, Leptodora) during summer in a mesotrophic lake. We hypothesized that due to differential size-selective predation risk by YOY fish and invertebrates, the large-sized and the small-sized Daphnia species would show different morphological responses to predation threats. We followed changes in two morphological traits (relative length of the tail spine in D. pulicaria and of the helmet in D. mendotae) among different periods during summer according to YOY fish and invertebrate predation. We defined four YOY fish predation periods based on the presence of YOY perch in the pelagic zone of the lake and the relative abundance of Daphnia preys in their gut contents, and two invertebrate predation periods based on exclusive or mutual occurrence of the invertebrate predators. The large-sized (D. pulicaria) and the small-sized (D. mendotae) species showed different morphological responses to YOY fish and invertebrate predators, respectively. The tail spine ratio of the juveniles and adults of D. pulicaria did not change in response to YOY fish predation or to invertebrate predation. A gradual increase in the helmet ratio was observed in the small-sized D. mendotae over the summer period. This change was related to the co-occurrence of the invertebrate predators (Chaoborus and Leptodora) and to YOY fish predation. The warmer temperature cannot be accounted for helmet elongation since it was constant across depths, and not related with the co-occurrence of D. mendotae and YOY perch. Guest editor: Piet Spaak Cladocera: Proceedings of the 7th International Symposium on Cladocera     

Biosynthesis of protease by Pseudomonas aeruginosa MN7 grown on fish substrate

Fish powders and fish protein hydrolysates (FPH) from sardinella (Sardinella aurita) were prepared and tested as growth media for alkaline protease production by Pseudomonas aeruginosa MN7. Cultivated in fish substrate as carbon source, the strain exhibited a slightly greater protease production (about 7800 U ml^–1) than that obtained with commercial peptones (about 7222 U ml^–1). Furthermore, P. aeruginosa MN7 produced the same amount of protease when cultivated in medium containing only fish substrate or that containing all ingredients, indicating that the strain can obtain its carbon and nitrogen requirements directly from whole fish proteins. Moreover, it was found that extensive hydrolysis of fish proteins did not increase protease formation. Protease production in media containing only FPH prepared by Alcalase was about 70% of those obtained with MN7 protease digest of fish protein or with meat-fish powder. These results indicate that sardinella substrates are an excellent carbon and nitrogen source for the growth of P. aeruginosa MN7 and the production of protease.     

Selective zooplankton predation by pre-adult roach (Rutilus rutilus): The size-selective hypothesis versus the visibility-selective hypothesis

Prey selection mechanisms of juveniles of the particulate feeding roach (Rutilus rutilus) were tested in bag experiments in lake Gjersjøn.Cladocerans and large developmental stages (cop. IV-ad.) of copepods were consumed in large numbers both in the bags added fish and in lake, while rotifers increased in numbers in bags added fish.The effect of fish predation on the small cladoceran Bosmina longirostris was tested by comparing morphological structures in fish-free bags and bags added fish.The results strongly support the visibility selective hypothesis, as eye-diameter became significantly smaller in bags added fish (P < 0.05, T-test), while body-size, length of antennae and mucro, frequency and number of eggs were slightly or unsignificantly correlated with presence or absence of fish.Further, while roach was found to consume large numbers of Bosmina longirostris, the far greater but nearly unpigmented Asplanchna priodonta was never consumed. Additionally, other data further supporting the visibility-selective hypothesis are discussed.     

Learned or innate production of acoustic signals in fishes: a test using a cyprinid

No information on the inheritance of the ability to produce sounds exists for fishes. In birds, which usually provide extensive post-hatching parental care, acoustic signals are learned in some species but are innate in others. Almost no fishes provide extensive post-hatching parental care and, consequently, the offspring have little opportunity to hear and learn sounds produced by the parents (usually the male in fishes); they may, however, be exposed to acoustic signals of conspecifics in the same habitat. We used a cyprinid, Codoma ornata, to test whether sound production is learned from the parents or whether it is innate. Fertilized eggs of this species were raised in isolation from adults. Upon maturity, these fish were tested for sound production in aggressive and reproductive contexts. Fish which had no contact with adults, and therefore no opportunity to hear the acoustic signals of their species, produced sounds that were similar to those produced by their parents, and they produced these in the same contexts. Significant differences were observed in dominant frequency for one context, with the smaller F₁ fish having signals of higher frequency than parental fish. Since no opportunity for learning existed, this provided evidence that the ability to produce sounds is innate in this minnow species.     

The predation impact of juvenile herring Clupea harengusand sprat Sprattus sprattus on estuarine zooplankton

The consumption of estuarine copepods by juvenile herring and sprat during estuarine residency was estimated using fish biomass data and daily rations calculated from two models of feeding in fish: a bioenergetic model and a gastric evacuation model. The bioenergetic model predicted daily rations that were, on average, three times higher than those estimated by a model based on field records of stomach contents. The biomass of herring and sprat in the estuary was negatively correlated with the daily ration suggesting that the clupeid fish populations were resource-limited. Copepod production decreased towards the winter and peaked in spring and summer. The relative importance of predation changed seasonally in function of the migration pattern of herring and sprat. In the spring and the summer, in situ production ofcopepod biomass was higher than the in situ consumption by fish. During the fall and the winter, consumption exceeded production. This suggests that top–down control exerted by marine pelagic fish may be an important force structuring estuarine copepod populations.     

Carbohydrate utilisation by microbial symbionts in the marine herbivorous fishes Odax cyanomelas and Crinodus lophodon

Carbohydrate uptake and catabolism by the gut microbiota of two species of temperate marine herbivorous fish were investigated using enzyme extracts prepared from microbial pellets. The fish studied were the herring cale Odax cyanomelas (Family Odacidae), which feeds on Ecklonia radiata, and the sea carp Crinodus lophodon (Family Aplodactylidae), which feeds primarily on red and green algae. Constitutive phosphoenolpyruvate phosphotransferase systems for glucose, galactose, fructose and mannitol were present in the microbiota of both fish. Hexokinase, fructokinase and mannitol dehydrogenase activities indicated that transport of the corresponding substrates may be coupled to permeases. Galactokinase activity was only detected in C. lophodon, as expected from its diet. Phosphofructokinase and pyruvate kinase activities were taken to indicate that carbohydrate metabolism proceeded via the fructose bisphosphate pathway. Differences in the transport and metabolism of the different monomers by the microbiota of O. cyanomelas and C. lophodon correlated strongly with predicted monomer availability in the gut of each species, suggesting that the microbiota are an integral component of digestion in these fish. The rates of production in adult fish of acetate, the major short-chain fatty acid, were estimated as 136 mol·h^-1 in O. cyanomelas and 166 mol·h^-1 in C. lophodon. These rates indicate that microbial fermentation is a potentially important source of energy for the host fish.Abbreviations AK acetate kinase - CTAB cetyl trimethylammonium bromide - FK fructokinase - F1P fructose 1-phosphate - F1PK fructose 1-phosphate kinase - F1-6BP Fructose 1,6-bisphosphate - F6P fructose 6-phosphate - GK galactokinase - Gal1P galactose 1-phosphate - G6P glucose 6-phosphate - HK hexokinase - MDH mannitol dehydrogenase - M1P mannitol 1-phosphate - M1-PDH mannitol 1-phosphate dehydrogenase - PEP phosphoenolpyruvate - PFK phosphofructokinase - PK pyruvate kinase - PTS phosphotransferase system - SCFA short-chain fatty acid(s) - TFA trifluoroacetic acid     

The effects of endogenous or exogenous catecholamines on blood respiratory status during acute hypoxia in rainbow trout (Oncorhynchus mykiss)

Summary An extracorporeal circulation of rainbow trout (Oncorhynchus mykiss) was utilized to continuously monitor the rapid and progressive effects of endogenous or exogenous catecholamines on blood respiratory/acid-base status, and to provide in vivo evidence for adrenergic retention of carbon dioxide (CO₂) in fish blood (cf. Wood and Perry 1985). Exposure of fish to severe aquatic hypoxia (final P _wO₂=40–60 torr; reached within 10–20 min) elicited an initial respiratory alkalosis resulting from hypoxia-induced hyperventilation. However, at a critical arterial oxygen tension (P _aO₂) between 15 and 25 torr, fish became agitated for approximately 5 s and a marked (0.2–0.4 pH unit) but transient arterial blood acidosis ensued. This response is characteristic of abrupt catecholamine mobilization into the circulation and subsequent adrenergic activation of red blood cell (RBC) Na⁺/H⁺ exchange (Fievet et al. 1987). Within approximately 1–2 min after the activation of RBC Na⁺/H⁺ exchange by endogenous catecholamines, there was a significant rise in arterial PCO₂ (P _aCO₂) whereas arterial PO₂ was unaltered; the elevation of P _aCO₂ could not be explained by changes in gill ventilation. Pre-treatment of fish with the -adrenoceptor antagonist phentolamine did not prevent the apparent catecholamine-mediated increase of P _aCO₂. Conversely, pre-treatment with the -adrenoceptor antagonist sotalol abolished both the activation of the RBC Na⁺/H⁺ antiporter and the associated rise in P _aCO₂, suggesting a causal relationship between the stimulation of RBC Na⁺/H⁺ exchange and the elevation of P _aCO₂. To more clearly establish that elevation of plasma catecholamine levels during severe hypoxia was indeed responsible for causing the elevation of P _aCO₂, fish were exposed to moderate hypoxia (final P _wO₂=60–80 torr) and then injected intraarterially with a bolus of adrenaline to elicit an estimated circulating level of 400 nmol·l^-1 immediately after the injection. This protocol activated RBC Na⁺/H⁺ exchange as indicated by abrupt changes in arterial pH (pHa). In all fish examined, P _aCO₂ increased after injection of exogenous adrenaline. The effects on P _aO₂ were inconsistent, although a reduction in this variable was the most frequent response. Gill ventilation frequency and amplitude were unaffected by exogenous adrenaline. Therefore, it is unlikely that ventilatory changes contributed to the consistently observed rise in P _aCO₂. Pretreatment of fish with sotalol did not alter the ventilatory response to adrenaline injection but did prevent the stimulation of RBC Na⁺/H⁺ exchange and the accompanying increases and decreases in P _aCO₂ and P _aO₂, respectively. These results suggest that adrenergic elevation of P _aCO₂, in addition to the frequently observed reduction of P _aO₂ are linked to activation of RBC Na⁺/H⁺ exchange. The physiological significance and the potential mechanisms underlying the changes in blood respiratory status after addition of endogenous or exogenous catecholamines to the circulation of hypoxic rainbow trout are discussed.Abbreviations P _aCO₂ arterial carbon dioxide tension - P _aO₂ arterial oxygen tension - P _da dorsal aortic pressure - pHa arterial pH - P _wO₂ water oxygen tension - RBC red blood cell - V _f breathing frequency     

Effect of temperature on the adsorption and one-step growth of the Nostoc virus N-1

This study was an attempt to observe the effects of temperature on adsorption and one-step growth of the virus N-1 infecting the nitrogen-fixing cyanobacterium Nostoc muscorum. Adsorption rate was found to be maximum at 40° C whereas no adsorption occurred at 10° C. The Q ₁₀ value was about 2.03 and the energy of activation, Ea was 16.3 kcal/ mole for the adsorption process. The development cycle of the virus was temperature sensitive. With increase in temperature, a gradual increase in inhibition of virus yield i.e. 8.33% at 30° C, 35.3% at 35° C and complete inhibition at 40° C was observed. Out of 7 h latent period, the early 4 h were temperature sensitive and heat treatment had a reversible inhibitory effect on virus development. The temperature treatment did not affect the rise period but burst-size was reduced.List of Abbreviattions PFU plaque-forming units - IM input multiplicity     

Life history responses of <Emphasis Type="Italic">Daphnia longispina</Emphasis> to mosquitofish (<Emphasis Type="Italic">Gambusia holbrooki</Emphasis>) and pumpkinseed (<Emphasis Type="Italic">Lepomis gibbosus</Emphasis>) kairomones

In the present study, the effect of chemical cues from two fish species (mosquitofish and pumpkinseed), at different concentrations, was tested in life history experiments with Daphnia longispina. The two fish species used represent the most abundant planktivores of many Mediterranean shallow lakes (SW Europe), where the indigenous fish communities have been replaced by such exotic assemblages. Results have shown a similar response of D. longispina to both fish species: kairomones stimulated daphnids to produce more offspring, which resulted in higher fitness (r), relatively to a fishless control. Fish presence also induced an earlier first reproduction, a smaller size at maturity of daphnids, and the production of smaller-sized neonates. Significant correlations with fish concentration (indirect measure of fish kairomone concentration) were found for size at maturity and neonate size, for both fish species. These results are in accordance to the “positive response” observed by other authors, which represents a defence mechanism to face losses caused by fish predators. The chemically mediated size reduction of mature females and neonates is an adaptive response to the size-selective predation exerted by fish. Pumpkinseed introduction is very recent in the lake of origin of the daphnids used in the experiments and its kairomone produced similar effects to mosquitofish in the life history of D. longispina. These results are contrary to the existence of a species-specific kairomone and support the hypothesis of a general fish kairomone. Guest editor: Piet Spaak Cladocera: Proceedings of the 7th International Symposium on Cladocera     

Preparation and testing of Sardinella protein hydrolysates as nitrogen source for extracellular lipase production by Rhizopus oryzae

Various fish protein hydrolysates (FPH) from sardinella (Sardinella aurita) were used as nitrogen sources for the production of extracellular lipase by the filamentous fungus Rhizopus oryzae. The best results were obtained with defatted meat–fish protein hydrolysates (DMFPH), indicating the presence in the lipid fraction of some constituents which may repress lipase synthesis. Furthermore, it was found that the extensive hydrolysis of fish proteins resulted in a higher lipase production. The use of 40 g DMFPH l^–1 for the growth of Rhizopus oryzae in medium R1 resulted in a lipase production of 394 U ml^–1, higher than the yield obtained with standard soy peptone as nitrogen source (373 U ml^–1). The most appropriate medium for the growth and the production of lipase is composed only of 24 g DMFPH l^–1 and 10 g glucose l^–1, indicating that the strain can obtain its nitrogen and salts requirements directly from fish substrate.     

Microalgae aquaculture feeds

Microalgae feeds are currently used in relatively small amounts in aquaculture, mainly for the production of larvae and juvenile shell- and finfish, as well as for raising the zooplankton required for feeding of juvenile animals. The blue-green algaSpirulina is used in substantial amounts (over 100 t y^–1) as a fish and shrimp feed, and even larger markets can be projected if production costs could be reduced. Another potential large-scale application of microalgae is the cultivation ofHaematococcus for the production of the carotenoid astaxanthin, which gives salmon flesh its reddish color. In the long-term microalgae biomass high in lipids (omega-3 fatty acids) may be developed as substitutes for fish oil-based aquaculture feeds. In shrimp ponds the indigenous algal blooms supply a part of the dietary requirements of the animals, but it is difficult to maximize algal productivities. A separate algal production system could feed the shrimps and minimize the need for added feed. Bivalves feed essentially exclusively on marine microalgae throughout their life cycle. The development of cultivation technologies for such microalgae would allow the onshore production of these animals, with greatly improved product quality and safety.This paper was presented at the Symposium on Applied Phycology at the Fourth International Phycological Congress, Duke University.