Behavioural and electrophysiological responses to F-prostaglandins, putative spawning pheromones, in three salmonid fishes

Behavioural and electro-olfactogram (EOG) responses to synthetic F-prostaglandins (PGFs) were recorded in the three salmonids: brown trout Salmo trutta , lake whitefish Coregonus clupeaformis and rainbow trout Oncorhynchus mykiss . Exposure to 10⁻⁸ M PGF_2α and 13, 14-dihydro-PGF_2α increased swimming activity in individually exposed brown trout in a flow-through tank. Digging and nest probing behaviours were further observed in brown trout females exposed to PGF_2α. Lake whitefish exposed to 10⁻⁸ M PGF_2α and 15-keto-PGF_2α also increased their locomotion. In rainbow trout, the absence of behavioural responses to PGFs correlates with a lack of olfactory sensitivity to these chemicals. PGFs triggered behavioural responses distinct from the feeding stimulant in brown trout. EOG measurements demonstrated that brown trout were most sensitive to PGF_2α, with a threshold concentration of 10⁻¹¹ M. Lake whitefish were most sensitive to both 15-keto-PGF_2α and 13, 14-dihydro-PGF_2α. Cross-adaptation and binary mixture experiments suggest that only one olfactory receptive mechanism is involved in PGFs detection. The behavioural and olfactory responses observed with exposure to PGF_2α and its metabolites suggest these compounds function as reproductive pheromones in brown trout and lake whitefish.     

Taste responses of the facial and glossopharyngeal nerves to amino acids in the rainbow trout

No significant differences were noted between responses of rainbow trout Oncorhynchus mykiss facial and glossopharyngeal nerves to 15 amino acids. Nine of these amino acids tested at 10⁻² M were stimulatory, whereas only two tested at 10⁻³ M were effective gustatory stimuli. For both nerve systems, ≤10⁻³ M L-proline was the most stimulatory amino acid, with an estimated threshold of 10⁻⁷ M; however, L-α-amino-β-guanidino-propionic acid (estimated threshold of 3×10⁻³ M), was the most potent compound at 10⁻² M. These results indicate that the same amino acids activate taste buds innervated by facial and glossopharyngeal nerves, respectively, and suggest that the same amino acids can be important in chemosensory feeding behaviour in the rainbow trout.     

Stimulatory effectiveness of amino acids on the olfactory response in an algivorous marine teleost, the rabbitfish Siganus fuscescens Houttuyn

The detection of threshold concentrations and relative stimulatory effectiveness (RSE) for 19 amino acids were studied by olfactory bulbar electroencephalogram (EEG) in the algivorous rabbitfish. The threshold concentrations for 19 amino acids ranged from 10⁻¹⁰ to 10⁻⁵ M. l -Alanine was the most effective amino acid, and the threshold concentration was estimated to be from 10⁻¹⁰ to 10⁻⁹M. These results were compared with the olfactory response in herbivorous and carnivorous fishes previously reported. The comparison of RSE for rabbitfish with those for other fishes showed correlation coefficients ranging from r = 0.97 to r = 0.50, but high similarities did not correspond with difference in feeding habits for herbivores and carnivores. The role of amino acids in fish chemosensory behaviour and the olfactory response of the rabbitfish was compared with the gustatory response reported previously. It was found that l -serine was the most potent stimulant in both chemoreceptors, l -proline and l -glutaminic acid were the most potent gustatory stimulants, and l -alanine, l -glutamine, l -arginine and l -lysine were the most potent olfactory stimulants.     

Cardiovascular responses of eggs, embryos and alevins of Atlantic salmon and rainbow trout to nitric oxide donors, sodium nitroprusside and isosorbide dinitrate, and inhibitors of nitric oxide synthase, N-nitro-l-arginine methyl ester and aminoguanidine, following short- and long-term exposure

Atlantic salmon embryos and alevins Salmo salar that had been exposed to isosorbide dinitrate (ISDN) for 4 weeks, on transfer to fresh water, showed an increase in heart rate. Unexposed embryos and alevins showed a decrease in heart rate following transfer to 100 μmol l⁻¹ ISDN for 4 h. This is in contrast to adult rainbow trout and higher vertebrates where tachycardia occurred in response to nitric oxide (NO) donors. The decreased heart rate in response to ISDN was inhibited by 2 mg 1⁻¹ methylene blue, indicating that NO activates cardiovascular events via guanylyl cyclase and cyclic guanidine monophosphate. Heart rate of rainbow trout alevins Oncorhynchus mykiss exposed to 100 μmol l⁻¹ aminoguanidine responded with a slowly developed but significant bradycardia over 10 min as did those reared in aminoguanidine for 4 weeks then transferred to fresh water. A potentiated increase in heart rate on exposure to the NO donor sodium nitroprusside (SNP), occurred within 1 min in salmon alevins reared in l -nitro-arginine methyl ester ( l -NAME) for 4 weeks, indicating up-regulation of NO receptors. The evidence for down-regulation of SNP-reared alevins exposed to l -NAME was less well defined. The results suggest that both salmonid embryos and alevins have a functional l -arginine-NO pathway and that NO has a physiological role in control of cardiovascular events.     

Gustatory responses to feeding‐ and non‐feeding‐stimulant chemicals, with an emphasis on amino acids, in rainbow trout

Specific receptor and fibre types of rainbow trout Oncorhynchus mykiss involved in the detection and discrimination of amino acids and a heterogeneous collection of compounds were investigated by recording the electrical activity of the maxillary branch of the facial nerve innervating taste buds inside the upper jaw. Proline (Pro), alanine (Ala), leucine (Leu), betaine (Bet) and 2‐amino‐3‐guanidinopropionic acid (Agp) were the major amino acids detected by the gustatory system. The two experimental approaches, concentration‐response curves and cross‐adaptations, showed that all amino acids were detected by three independent receptor types: Pro ‐, Agp/Bet ‐ and Leu ‐receptors. Bile acids, the most potent stimulants recorded, were detected by a single receptor type independent of those for amino acids, with threshold concentrations of 10⁻¹² M. Strychnine, quinine and tetrodotoxin may have partially shared a single receptor mechanism. The gustatory sensibility narrowly tuned towards the amino acid spectrum compared to those for a diverse array of non‐feeding stimulant chemicals, combined with feeding behaviour triggered primarily by vision and olfaction, suggest that in rainbow trout, and possibly other salmonid species, gustatory chemical cues, in addition to food finding and intake, play an important role in detecting poisonous prey and substances.     

STIMULATORY EFFECTS OF SUBSTANCE P ON NEURITE EXTENSION AND CYCLIC AMP LEVELS IN CULTURED NEUROBLASTOMA CELLS

Abstract— Synthetic substance P initially increased cyclic AMP levels and subsequently induced neurite extension in cultured neuroblastoma N 18 cells. The magnitude of these effects depended on the concentration of fetal calf serum (FCS) in the culture medium, being more evident in the presence of a lower (0.1%) concentration of FCS.
In Eagle's medium containing 0.1% FCS, low concentrations of substance P (10⁻⁷-10⁻⁵ M) increased cyclic AMP levels and stimulated neurite extension.
In Eagle's medium containing 5%FCS, both substance P at concentrations of 10⁻⁵-10⁻³M and dibutyryl cyclic AMP at concentrations of 10⁻⁴-10⁻²M increased cyclic AMP levels and stimulated neurite extension. The activities of acetylcholinesterase, (Na⁺+ K⁺)-, HCO₃⁻ and Mg²⁺ -stimulated-ATPase were also increased. Cell growth was inhibited.
Substance P at concentrations of 10-⁷-10⁻⁵M also stimulated the adenylate cyclase activity of a particulate fraction of N 18 in a concentration-dependent manner.     

Feeding behaviour in some teleosts is triggered by single amino acids primarily through olfaction

The present study investigated behavioural responses of naïve rainbow trout Oncorhynchus mykiss , lake charr Salvelinus namaycush , lake whitefish Coregonus clupeaformis and goldfish Carassius auratus to food extracts as well as selected amino acids (at 10⁻⁶ M) and related chemicals in a flow‐through behaviour trough. Cysteine, the most potent olfactory stimulating amino acid determined electrophysiologically, enhanced locomotor activity, an initial arousal behaviour, in all species examined, in exactly the same fashion as did food extracts. The increased locomotor activity was followed by distinct species‐specific search behavioural patterns: 1) bottom searching in rainbow trout; 2) surfacing and jumping in lake charr; 3) exploratory behaviour against the trough window in lake whitefish; 4) gravel pecking in goldfish. Olfactory‐stimulating neutral amino acids alanine and serine and gustatory‐stimulating imino acid proline were effective, to varying degrees, in eliciting feeding behaviour. Arginine and glutamate suppressed the locomotor activity in rainbow trout, but triggered feeding behaviour in the other species. Agmatine, a decarboxylated arginine, stimulated feeding behaviour in all species tested, except rainbow trout. The observed behavioural responses were in close accordance with the electrophysiological threshold and specificity. Together, the present data demonstrate that single amino acids and closely related chemicals initiate feeding behaviour primarily by olfaction, and interchangeably and complemented by gustation, in naive fishes.     

Fish tongues–surface fine structures and ecological considerations

The surface structures of the tongues of Dissostichus mawsoni, Trematomus bemacchii, f. borchgrevinki (Nototheniidae), Stemoptyx diaphana (Sternoptychidae), Diretmus sp. (Diretmidae), Cataetyx memorabilis (Ophidiidae), Photoblepharon palpebratus (Anomalopidae), Carapus mourlani (Carapidae) and Salmo gairdneri (Salmonidae) were investigated with the scanning electron microscope. The aim of the study was to test the theory that the degree to which gustatory receptors are developed morphologically in different species of fishes is related to the environment, and in particular to the variey of food present in the latter. It was found that in the two mesopelagic species, S. diaphana and Diretmus sp., taste receptors were poorly developed, but in the benthic deep-water fish C. memorabilii , which lives off South West Africa in a biologically rich environment, well-developed taste receptors occurred in large numbers. The three Antarctic species studied had similar tongue surface structures with rather well developed papillae of a moderate density. Photoblepharon palpebratus , in spite of its small tongue, possessed a large number of taste buds on elevated ridges; Carapus mourlani , however, had only very few obvious receptors and is thought to be a poor taster. Undoubtedly the highest degree of taste receptor development is found in the freshwater trout Salmo gairdneri. The results support the hypothesis that taste receptor development and the variety of food types preseni in a fish's environment are correlated.     

Effects of egg composition and prey density on the larval growth and survival of lake whitefish (Coregonus clupeaformis Mitchill)

Eggs were collected from two stocks of lake whitefish, Coregonus clupeaformis , in Lakes Michigan and Huron to assess the effect of egg composition and prey density on larval growth and survival. Egg composition parameters including wet weight (mg egg⁻¹), dry weight (mg egg⁻¹), percent water, total caloric content (cal egg⁻¹), caloric density (cal egg⁻¹), percent lipid content, and total lipidcontent (mg egg⁻¹) were measured. Fish hatched from six parental females in each stock were fed one of four rations (0, 18, 24, 50 brine shrimp larva⁻¹ day⁻¹) after yolk sac absorption. Length at hatch, endogenous growth, exogenous growth, and survival were measured during a 42-day laboratory experiment. Length at hatch of larvae was positively related to egg caloric content ( r ²=0.780). Endogenous growth for lake whitefish larvae was positively related to percent lipid content ( r ²=0.896) and total egg lipid content ( r ²=0.876) of parental females. Exogenous growth and survival of larval lake whitefish was positively related to prey availability. Larval fish growth was accurately modelled ( r ²=0.973) as a function of prey abundance using a threshold-corrected hyperbolic equation. These results indicate that both egg composition and prey availability have the potential to influence the growth and survival dynamics of larval lake whitefish significantly.     

The relative abundance of brown trout in acidic softwater lakes in relation to water quality in tributary streams

The influence of the water quality of tributary streams on the relative abundance in benthic gillnet catches (catch per unit effort, cpue) of allopatric brown Salmo trutta was assessed in associated acidic, softwater lakes. The study was carried out over 6 years (1989–1994) in 15 lakes located at altitudes between 230–715 m a.s.l. in two Norwegian catchments. The water quality of the main inlets and outlets varied little, as indicated by their of pH range (4·93–5·51) and calcium concentrations (0·19–0·44 mg 1⁻¹), but varied more with respect to concentrations of inorganic, monomeric aluminium (7·0–41·0 μg l⁻¹). Most of the lakes were also fed by secondary streams with better water quality: a maximum pH of 6·56, calcium levels of up to 0·74mg 1⁻¹, and inorganic aluminium levels as low as l·0 μg 1⁻¹. The cpue was inversely correlated with lake altitude ( r ²=0·50), and thus was adjusted to a mean altitude. The calcium concentration in the richest secondary stream to each lake, its richness judged on the basis of its acid-neutralizing capacity, had the highest predictive power of the variability in cpue ( r ² = 0·49).The calcium content in the other secondary streams or in the main inlets and outlets did not correlate with cpue. Alkalinity in the main outlets correlated to some extent with cpue ( r ² = 0·27). It is suggested that secondary streams with good water quality provide important refuges for the recruitment of brown trout in acidic softwater lakes.     

Prediction, from temperature, of eyeing, hatching and 'swim-up' times for salmonid embryos

SUMMARY. 1. The literature contains a number of curves relating time (days) required for median hatch (=D₂) to water temperature (T,°C) for the eggs of several salmonid fishes. There are relatively few data on the relationships between time to median eyed (= D₁), days) or time to median swim-up (= D₃, days) and temperature.
2. From published data, over most of the range 0-9.5°C, approximate relationships are D,=0.5D₂ for Atlantic salmon (Salmo salar L.) and D3 = 1.7D₂ for eight species of Salmo and Oncorhynchus.
3. Field and hatchery tests suggest that these are useful empirical models for approximate prediction of D₁ and D₃ from D₂ for most salmonids.     

Behavioural responses of fishes to chemical cues: results from a new bioassay

A new behavioural bioassay for determining the locomotor response by fish to chemical cues is described. The assay combines a behavioural trough, capable of delivering up to 10 discrete plumes of treated water, together with a digital method for quantifying the movements of single fish. Sample responses of two whitefish, Coregonus clupeaformis , one to an attractant (a food extract) and one to a repellant (quinine monohydrochloride), are provided. Their locomotor responses are analyzed in terms of such parameters as total distance travelled, average velocity, number of turns, average excursion distance, and the number of times specific boundaries are crossed. The assay's sensitivity is demonstrated through concentration-response curves by Arctic char, Salvelinus alpinus , to the bile acid, taurocholic acid, and the amino acids, L-serine and L-alanine. Behavioural response threshold concentrations for these three substances were estimated to be 10⁻¹⁵, 10⁻ⁿ, and 10⁻⁹ m, respectively.     

Effect of rigidity of gel medium on benzyladenine-induced adventitious bud formation and vitrification in vitro in Picea abies

Observations on the effects of different degrees of rigidity of both an agar (Tayio) and a non-agar (Gelrite) gel on the uptake of radiolabelled N⁶-benzyladenine (¹⁴C-BA) were also extended to mode of application and positioning of the explant. Regression analysis showed a highly significant inverse correlation between ¹⁴C-BA accumulation and degree of gel stiffness. Significantly greater numbers of adventitious buds per explant were induced at low to medium levels of rigidity (2.5–10 g Tayio 1⁻¹, 1–5 g Gelrite 1⁻¹); this advantage was almost completely nullified at the lower levels (2.5 and 5.0 g Tayio 1⁻¹, 1 and 1.5 g Gelrite 1⁻¹) as a result of the high incidence of vitrification. In addition to turgor distension, vitrified buds displayed cellular damage. Explants with their cotyledons flattened onto the agar surface accumulated less ¹⁴C-BA after 96 h than upright explants, but produced greater numbers of adventitious buds, pseudobuds and phylloids. It was suggested that BA was taken up only by "target" cells, presumably the differentiating subsidiary cells of those stomatal complexes in surface contact with the medium. Pulse treatments of relatively short durations (2 h) with optimal concentrations of BA (ca 125 μ M ), followed by subculturing on hormone-free media gelled with 10 g agar 1⁻¹, produced a satisfactory balance between yield and competence of adventitiously-induced buds.     

Behavioural responses of salmonid fry to low amino acid concentrations

The behaviour of start feed brown trout, Salmo trutla , and Atlantic salmon, Salmo salar , fry was observed in response to 5, 50 and 500 nM concentrations of L-alanine, _L-proline, L-arginine and glycine. In addition, salmon fry were tested with dilute concentrations of shrimp extract. Five behaviour patterns (snap, yawn, dart, twitch movements and active swimming) were shown in response to all amino acid and shrimp extract concentrations. Snapping, darting and active swimming increased in both species as a function of amino acid concentration, and in the salmon fry as a function of increasing shrimp extract concentration. Otherwise, the salmon showed twice as much yawning and more twitch movements than the trout, but the trout showed more active swimming than the salmon. Both species showed an increase in activity in response to L-proline at 5 nM, and the salmon also responded to L-arginine and glycine at this concentration. Both species first responded to L-alanine at 50 nM, but the trout did not respond to glycine until a concentration of 500 nM was presented. The salmon fry responded to shrimp extract at c. 10 ¹⁴g l^-1, but no differences in their activity were observed in response to concentrations between c. 10^-12 and 10^-6gl^-1.     

DOMOIC ACID PRODUCTION IN NITZSCHIA SP. (BACILLARIOPHYCEAE) ISOLATED FROM A SHRIMP-CULTURE POND IN DO SON, VIETNAM

Domoic acid (DA), a neuroexcitatory amino acid, was detected in batch culture of the newly recognized species Nitzschia navis-varingica Lundholm et Moestrup . The production of DA by this diatom was confirmed by electrospray ionization mass spectrometry. The diatom was collected from a shrimp-culture pond in Do Son, Vietnam. The production of DA (1.7 pg·cell ^{− 1}) is within the levels reported for Pseudo-nitzschia multiseries (Hasle) Hasle. The DA production started during the late exponential growth phase and reached a maximum during the early stationary growth phase. Maximum DA levels in the axenic culture decreased to about half that of the nonaxenic culture (0.9 pg·cell ^{− 1} vs. 1.7 pg·cell ^{− 1}), suggesting that DA production by the new species is influenced by bacteria.     

Juvenile salmonid production in a Lake Erie nursery stream

Normandale Creek (2531m²) provides spawning and nursery grounds for lake-run rainbow trout, Salmo gairdneri Richardson, brown trout, Salmo frutta L. , and coho salmon, Oncorhynchus kisutch (Walbaum). Upstream movements are significantly correlated with peak stream discharge (P < 0.05). In 1973–1974, 59 adult salmonids constructed 86 nests of which 60% were disturbed by re-use or sand deposition. From a calculated deposition of 90 400 ova, 483 juveniles, 6 cm in fork length ( f.l. ), were resident in the stream when estimates by electrofishing first became valid. Estimated within-stream (i.e. all sections combined) monthly densities of wild juveniles >6 cm f.l. ranged from six to 22 fish (100 m²)⁻¹, and biomass from 1.3 to 3.1 gm⁻². The highest within-section biomass was 8.3 gm⁻² in November. Additions of hatchery-reared rainbow trout temporarily increased monthly within-stream juvenile density up to 23 fish (100 m²)⁻¹, and biomass to 8.2 g m⁻². Density and biomass were positively correlated (P < 0.05) with both instream and bank cover, and biomass negatively correlated (P < 0.05) with gradient. The importance of substratum and flow characteristics on juvenile density and biomass was strongly indicated although not statistically significant. Annual within-section production of juveniles > 6 cm f.l. ranged from 4.79 to 5.93 g m⁻² year⁻¹ in Sections III and IV, respectively. Inclusion of calculated probable numbers of fish < 6 cm f.l. increased within-section production from 5.93 to 18.65 g m⁻² year^−l in upstream Section IV.     

Effect of rearing density on thyroid and interrenal gland activity and plasma and hepatic metabolite levels in rainbow trout, Salmo gairdneri Richardson

There were significant inverse correlations between rearing density of rainbow trout, Salmo gairdneri Richardson, and final body weight, plasma L-thyroxine (T₄), trüodo-L-tryronine (T₃), cortisol and protein concentrations, plasma T₄/T₃ ratios and thyroid epithelial cell height. In addition, hepatosomatic indices and plasma free fatty acid concentrations were higher in fish reared at low (134 g 1⁻¹) density compared with groups reared at medium (210g1⁻¹) or high density (299g 1⁻¹), and the post-feeding (3.5-4h) elevation in plasma glucose and triglyceride levels evident in trout maintained at low rearing density was not found in those fish reared at higher densities. There were no significant effects of rearing density on hematocrit, carcass composition, hepatic glycogen and lipid levels and interregnal nucleus size.     

Effect of CO2-enrichnient on seedling physiology and growth of two tropical tree species

Seedlings of two tree species from the Atlantic lowlands of Costa Rica, Ochroma la-gopus Swartz, a fast-growing pioneer species, and Pentaclethra macroloba (Willd.) Kuntze, a slower-growing climax species, were grown under enriched atmospheric CO₂ in controlled environment chambers. Carbon dioxide concentrations were maintained at 350 and 675 μl 1⁻¹ under photosynthetic photon flux densities of 500 μol m⁻² s⁻¹ and temperatures of 26°C day and 20°C night. Total biomass of both species increased significantly in the elevated CO₂ treatment; the increase in biomass was greatest for the pioneer species, O. lagopus . Both species had greater leaf areas and specific leaf weights with increased atmospheric CO₂. However, the ratio of non-pho-tosynthetic tissue to leaf area also increased in both species leading to decreased leaf area ratios. Plants of both species grown at 675 μl 1⁻¹ CO₂ had lower chlorophyll contents and photosynthesis on a leaf area basis than those grown at 350 μl 1⁻¹. Reductions in net photosynthesis occurred despite increased internal CO₂ concentrations in the CO₂-enriched treatment. Stomatal conductances of both species decreased with CO₂-enrichment resulting in significant increases in water use efficiency.     

Resistance to cadmium by pretreated rainbow trout alevins

A toxicity test with cadmium concentrations ranging from 0·1 to 100·0 mg Cd 1⁻¹ was used to assess the effect of cadmium pretreatment on rainbow trout ( Salmo gairdneri Richardson) alevins. The median period of survival for fish pretreated at 0·01 mg Cd 1⁻¹ was found to be increased at test concentrations up to 10mg Cd 1⁻¹ compared with alevins pretreated with dilution water. However, at concentrations above 10mg Cd 1⁻¹ pretreatment at 0·01 mg Cd 1⁻¹ reduced the median period of survival.     

Temperature effects on anaesthesia with clove oil in six temperate‐zone fishes

The potential use of clove oil (eugenol) as an anaesthetic for Atlantic salmon Salmo salar , brown trout Salmo trutta , rainbow trout Oncorhynchus mykiss , whitefish Coregonus lavaretus , perch Perca fluviatilis and roach Rutilus rutilus was examined at 5, 10, 15 and 20° C using three anaesthetic concentrations (varying from 20 to 200 mg l⁻¹) at each temperature. Substantial species differences in sensitivity to clove oil were observed, even amongst congeners, and there may be some disadvantages (slow recovery and possibly mortality) with using clove oil for 0+ year whitefish and at low temperatures for perch and roach.