Effects of modulatory agents on neurally-mediated responses of trout intestinal smooth musclein vitro

Mediators and mechanisms responsible for the inhibitory modulation of trout intestinal smooth muscle were examined using a series of putative mediators and substances known to modulate neurotransmission in mammalian systems. Frequency response relationships to transmural stimulation and concentration response relationships to 5-hydroxytryptamine, carbachol, and substance P were established on paired segments of rainbow trout intestinein vitro in the presence and absence of putative modulatory agents. Modulation of neurally-mediated contractions of trout intestine was achieved with dibutyryl cyclic AMP and forskolin, agents that increase intracellular levels of cyclic AMP. The effect appears to be at the level of the smooth muscle, since the adenylate cyclase activator, forskolin, inhibited muscarinic and serotoninergic contractions as well as transmurally stimulated contractions. Substance P-induced contractions were unaffected by forskolin. The endogenous agonists/neurotransmitters which would increase cyclic AMP levels in rainbow trout intestinal smooth muscle are as yet unknown. The effects do not appear to be modulated by vasoactive intestinal peptide (VIP), calcitonin, calcitonin gene-related peptide (CGRP), or agents that activate -adrenoceptors. Prostaglandin E₂ (PGE₂) and ₂-adrenergenic agonists are possible agents which will decrease contractility of the smooth muscle. They were only active in the proximal intestine and on transmurally stimulated contractions. The effects of both PGE₂ and ₂-agonists appear to be prejunctional, decreasing release of contractile neurotransmitters in the enteric nervous system.     

Changes in smooth muscle contractility of rainbow trout (Oncorhynchus mykiss Walbaum) intestine during acclimation to altered temperature

The effects of altered water temperature in vivo on in vitro smooth muscle contractility of rainbow trout intestine were investigated. Temperature has a significant effect on receptor-mediated intestinal smooth muscle contractility in the rainbow trout. The efficacy of 5-HT, carbachol, and transmural stimulation increased with temperatures above 10°C, with an optimal increase at 15°C. There was also a modest increase in the potency of 5-HT and carbachol within 2 days of establishing trout at 20°C. By day 8, most of these changes had either stabilized or were returning to control values, suggesting that acclimation changes in membranes and enzyme activities were taking effect. However, the contractile responses to carbachol and transmural stimulation were still increasing at this time. This may imply that the muscarinic receptors are more resistant to membrane acclimation changes and may take longer to adapt. Because these experiments were controlled for handling stress and seasonal changes that affect contractility, we have been able to demonstrate some early changes in smooth muscle contractility that occur during acclimation to altered temperature.     

Effects of pH on contractility of rainbow trout (Oncorhynchus mykiss) intestinal muscle in vitro

Extreme acid-base imbalances in salmonids have been shown to impact on gastrointestinal motility. Changes in pH are known to affect cardiac and skeletal muscle in fish and mammalian intestinal smooth muscle preparations. This study was conducted to determine if the contractility of rainbow trout intestinal muscle was sensitive to pH fluctuations and to begin the investigation into the mechanisms by which pH affected contractility. Isolated duplicate or triplicate segments of rainbow trout intestine were suspended in organ baths containing modified Krebs-Henseleit solution. The pH was adjusted by varying the concentration of CO₂ aerating the solution and with HCl or lactic acid. Contractility was determined as response to the administration of 5-hydroxytryptamine (5-HT, serotonin), KCl, and transmural stimulation within the pH range of 6.3 to 8.5. Optimum pH for proximal segments was 7.85 while the range was much wider for distal segments. Decreasing the pH with CO₂ resulted in a greater inhibition of smooth muscle contractility than when the pH was decreased in 100% O₂ with either HCl or lactic acid, particularly when the tissues are stimulated electrically. This effect was ameliorated as the fish attained sexual maturity, although the mechanisms involved were not clear. The Cl^-/HCO ₃ ^- exchanger appeared to be involved in recovery from acidotic stress, particularly at pH levels below 6.5. The role played by the Na⁺/H⁺ exchanger is still unclear. Although contractility was inhibited in the presence of the amiloride analog EIPA (5-(N-ethyl-N-isopropyl)amiloride), whether the inhibition was due to blockage of ion transport or to cytosolic effects is yet to be determined.     

Muscarinic M3-like receptors, cyclic AMP and L-type calcium channels are involved in the contractile response to cholinergic agents in gut smooth muscle of the rainbow trout, Oncorhynchus mykiss

Cholinergic signalling in mammalian gut smooth muscle usually involves M3 muscarinic receptors for direct contraction via phospholipase C activation and M2 muscarinic receptors to reduce cyclic AMP levels. However, the proportion of receptor subtypes and second messengers involved varies among tissues and animals and studies in non-mammalian species will provide information on the conservation of pathways and consequently on their importance for signal transduction. In the present study we investigated receptor subtypes, involvement of calcium, phospholipase C and cyclic AMP in the cholinergic contraction of the rainbow trout gut. Intestinal and gastric smooth muscle strip preparations, with the mucosa removed, were used in functional studies, and homogenised strips were used for measurements of cyclic AMP. Calcium-free medium, the L-type calcium-channel inhibitor verapamil, the cyclic AMP-phosphodiesterase inhibitor IBMX + isoprenaline, and the M3-antagonist 4-DAMP methiodide all caused a partial or marked reduction of the response to cholinergic agonists. Neomycin, an inhibitor of phospholipase C, and SKF96365, an inhibitor of receptor-operated calcium channels, had no effect. Carbachol (0.1 mM) reduced the levels of cyclic AMP transiently. It is concluded that the cholinergic signal transduction in rainbow trout gut smooth muscle involves (1) binding to M3-like receptors, (2) a transient reduction in cyclic AMP levels, (3) influx of extracellular calcium, in part through L-type calcium-channels and (4) no involvement of phospholipase C.     

Immunohistochemical detection of neuromodulators in the intestine of Salmo trutta L. naturally infected with Cyathocephalus truncatus Pallas (Cestoda)

Pathological and immunohistochemical investigations were carried out on the middle intestine of uninfected and parasitized brown trout, Salmo trutta L., from the Ceresone Channel in North Italy. Eighty-six brown trout were sampled by electrofishing, and 32 (37.2%) were infected with Cyathocephalus truncatus Pallas, 1781 (Cestoda). The intensity of infection ranged from 1 to 82 parasites per host and the most infected segments were the anterior (near the pyloric caeca region) and the central part of the middle intestine. Immunohistochemical tests were applied on sections of intestinal tissue of healthy and infected fish, and the presence of substance P (SP), calcitonin gene related peptide (CGRP), met-enkephalin, vasoactive intestinal peptide (VIP), neuropeptide Y (NPY) and serotonin (5-HT) was documented. Endocrine epithelial cells of the tunica mucosa were positive to SP-, CGRP-, met-enkephalin-, and NPY-like peptides and 5-HT antisera; moreover, a higher number of these cells were recorded in the intestine of infected trout in comparison to uninfected fish. In addition, in parasitized S. trutta , SP-like and 5-HT immunoreactivities were found in likely immuno-related cells of the tunica propria-submucosa. Nerve cell bodies and terminals in the myenteric plexus were immunoreactive to almost all the tested peptides and 5-HT antisera. These data provide evidence for the role of the neuroendocrine system of S. trutta in the modulation of inflammatory responses to C. truncatus . Results are discussed with respect to a peptidergic involvement and host immune response to an intestinal tapeworm.     

Effects of vasoactive intestinal polypeptide,substance P, 5-hydroxytryptamine,met-enkephalin and neurotensin on the swimbladder smooth muscle of two teleost species,Gadus morhua andAnguilla anguilla

The effects of vasoactive intestinal polypeptide (VIP), substance P (SP), 5-hydroxytryptamine (5-HT), neurotensin (NT) and met-enkephalin (mEnk) on the smooth muscles of the teleost swimbladder were studied in two teleost species, the Atlantic cod (Gadus morhua) and the eel (Anguilla anguilla). The study was made on isolated strip preparations of the muscularis mucosae, using putative transmitters corresponding to the immunoreactive materials that have previously been localized by immunohistochemical methods in nerves or endocrine/paracrine cells of the teleost swimbladder and/or the gastrointestinal canal. VIP was relaxatory on both cod and eel swimbladder smooth muscle, SP and 5-HT were constrictory in both species, and mEnk was excitatory on the eel swimbladder smooth muscle. Clear effects of these agents were usually seen at a concentration ofca. 10 nM in cumulative concentration/effect experiments. NT had no effect in either species. In the eel, the effects on the pneumatic duct were generally greater than on the swimbladder proper. The study indicates that the 5-HT and peptides previously observed by immunohistochemistry have physiological functions in the swimbladder.     

Branchial cobalt uptake in the carp, Cyprinus carpio: Effect of calcium channelblockers and calcium injection

Using tracer-uptake studies we investigated whether the branchial uptakeof cobalt occurs via selective, inhibitable, calcium uptake routes.Modulation of the calcium transport system was performed using generalcalcium channel blockers (Cd²⁺, La³⁺,Mg²⁺), a voltage-dependent calcium channel blocker (diltiazem)and an intraperitoneal CaCl2 injection. Adding Cd²⁺ orMg²⁺ to thewater or injecting the fish with theCa²⁺-solution resulted in decreased Co²⁺ andCa²⁺ uptake rates. Addition of La³⁺ reducedCa²⁺ and Co²⁺ uptake rates except forCo²⁺ uptake in the gills. Diltiazem displayed only aninconsistent, concentration-independent blocking of the uptake of bothCo²⁺ and Ca²⁺. All inhibitors which inhibitCa²⁺ uptake also inhibit Co²⁺ uptake. However, aclear difference in the degree of inhibition of uptake of both elements isobserved. Two possible explanations are suggested (1) Co²⁺ andCa²⁺ are both transported through the gills via the sametransport system or (2) the calcium transport system is not the only systeminvolved in the transport of Co²⁺.     

The effects of acute temperature change on smooth muscle contractility of rainbow trout (Oncorhynchus mykiss Walbaum) intestine

The effects of altered temperature in vivo on in vitro smooth muscle contractility of rainbow trout intestine were investigated. Initial analysis of the data revealed a seasonal variation in the maximal tension of intestinal smooth muscle attainable with 5-hydroxytryptamine (serotonin), carbachol, KCl, and transmural stimulation in vitro. Peaks occurred in spring and troughs in autumn. There was no seasonal cycling of the potency of the stimulants. All data regarding the efficacy of the stimulants were subsequently corrected for seasonal variation. The response of smooth muscle depends on the temperature of the water in which the fish are placed (2°C−20°C). There was a marked linear increase in efficacy and a slight increase in potency of the stimulants with increasing temperature. Changes in responsiveness of the intestinal smooth muscle occurred within 30 min of moving the fish between tanks. Smooth muscle reactivity returned to pretreatment values by 48h. Any changes in responsiveness with regards to time were unlikely to be as a consequence of water temperature, but may have been a result of handling stress.     

Characterization and expression of SLO1 potassium channels during spermatogenesis in Eriocheir sinensis

SLO1 potassium channels are pivotal to many aspects of spermatogenic cell. Experiments were conducted to assess physiology and function of SLO1 potassium channels in different developmental stages of spermatogenic cell in Eriocheir sinensis. First, the expression of SLO1 protein was examined via Western blot, RT‐PCR, immunohistochemistry and immunofluorescence assays. The results showed that the expression of the SLO1 protein was not uniform in the spermatogenic cells of E. sinensis. Second, whole‐cell patch clamp technique was used to record the potassium current of spermatogenic cells and to analyse the electrophysiological characteristics of the potassium channels with the aid of inhibitors. It is proved that the potassium current in E. sinensis germ cells is associated with intracellular Ca²⁺, and the calcium‐activated potassium channel mediated by SLO1 protein is mainly large‐conductance Ca²⁺‐activated K⁺ channels (BK_Ca). Based on these researches, the cDNA of SLO1 from testis was cloned and sequenced. The SLO1 protein contained domains bound to calcium ions, and the spatial structure formed by its tetramers constituted potassium channels. Phylogenetic analysis revealed that SLO1 was much closer to Scylla paramamosain than other examined species. Finally, iberiotoxin (IbTX) and CdCl₂ were used to inhibit the acrosome reaction (AR) induced by A23187 and to explore the role of SLO1 potassium channels in the AR of E. sinensis. The experimental results showed that SLO1 potassium channels were expressed in E. sinensis spermatogenic cells and played an important role in the AR of crab sperm (SP).     

Estradiol-17β-induced calcium uptake and resorption in juvenile rainbow trout,Oncorhynchus mykiss

Juvenile rainbow trout, Oncorhynchus mykiss, were injected with estradiol-17β (E₂) in order to study the source of extra calcium needed during vitellogenesis. E₂-treatment increased the calcium uptake from the external medium as well as calcium mobilization from muscle and scale. Judged by the increase in plasma protein-bound calcium levels, the E₂-induced increase in calcium uptake is an apparent over-mobilization of calcium, i.e., the calcium uptake of the fish is in excess of what is found bound to plasma proteins. As the calcium excretion and calcium space (calculated from free plasma calcium levels) were unaffected, the excess calcium is suggested to be incorporated into internal calcium stores. This implies that the systems regulating vitellogenesis and calcium balance are integrated on the mechanistic or endocrine level, and that E₂ causes calcium mobilization of a magnitude geared to the needs of the sexually maturing female.     

Control of gallbladder motility in the rainbow trout,Salmo gairdneri

Immunohistochemistry revealed nerves containing VIP-like and 5-HT-like material in both gallbladder wall and bile duct of the rainbow trout, while endocrine cells containing gastrin/CCK-like and substance P-like material were present in the mucosa of the bile duct and the duodenum. Fluorescence histochemistry showed adrenergic nerves close to the muscle layer of the gallbladder.Sulphated CCK8, caerulein and non-sulphated CCK8 (in this order of potency), 5-HT and acetylcholine were excitatory on isolated strip preparations, while VIP and adrenergic agonists were inhibitory. The adrenergic drugs were probably actingvia a beta-adrenergic receptor, while the effects of 5-HT and cholinergic drugs were antagonized by methysergide and atropine respectively.Electrical stimulation of the gallbladder nervesin situ failed to show any effect or under certain conditions induced a rebound effect.It is concluded that the motility control of the rainbow trout gallbladder may involve an inhibitory innervation by adrenergic and possibly VIP-releasing nerves, while 5-HT, acetylcholine and a CCK-like substance may be involved in the excitatory control.     

Effect of feeding and fasting on plasma tryptophan and tryptophan to large neutral amino acid ratio,and on brain serotonin turnover in rainbow trout,Oncorhynchus mykiss

Two time-course experiments were conducted to determine the effect of feeding and fasting on the plasma ratio of tryptophan (trp) to the large neutral amino acids (LNAA), (trp/LNAA ratio) and brain serotonin (5-HT) turnover in rainbow trout,Oncorhynchus mykiss. Trout were fasted overnight or for 3 days and were then either fed or continued to be fasted for up to a further 3 days. Changes in plasma trp, plasma trp/LNAA ratio, brain trp, brain 5-HT, brain 5-hydroxyindoleacetic acid (5-HIAA) and brain 5-HIAA/5-HT ratio were determined over time. Feeding decreased the plasma trp/LNAA ratio, brain trp and the brain 5-HIAA/5-HT ratio. In addition, in fish sampled over 3 days, there appeared to be a rhythm in plasma trp and the brain 5-HIAA/5-HT ratio which was independent of feeding. These results indicate that in rainbow trout, feeding is a sufficient physiological event to decrease brain 5-HT turnover. Furthermore, feeding-independent changes in the brain 5-HIAA/5-HT ratio, which were evident in fasted fish sampled over 3 days, also suggest an additional, non-feeding-related modulator(s) of brain 5-HT turnover in rainbow trout.     

Calcium currents and exocytosis in single isolated pars intermedia cells from tilapia (Oreochromis mossambicus)

The role of calcium influx through voltage-dependent calcium ion channels in the exocytotic response of single isolated pars intermedia cells from the teleost tilapia (Oreochromis mossambicus) was investigated by means of the whole-cell patch-clamp technique and high resolution electrical measurements. Calcium currents differed from barium currents in several ways: the peak I_Ca was smaller, the current-voltage relationship for calcium attained its maximum at +10 mV instead of 0 mV, and the inward calcium current inactivated more rapidly. Electron micrographs showed that pars intermedia cells possess dense, encored vesicles with an average diameter of 140 nm. Influx of calcium resulted in an increased cell membrane capacitance (C_m) after the depolarizing period, indicating a fast exocytotic response. Comparison with late recordings revealed the presence of a transient in C_m, presumably attributable to movement of ion channel gates. The average increase in C_m was 13.4 fF, suggesting the fusion of at least 23 vesicles with the plasma membrane during the depolarizing pulse. In contrast to calcium, barium did not support significant exocytosis. We conclude that calcium entry through voltage-dependent calcium channels rapidly leads to the exocytosis of secretory vesicles from tilapia pars intermedia cells.     

Effects of dietary supplements on the availability of minerals in fish meal; preliminary observations

Preliminary studies were conducted to determine if several feed supplements with the potential to improve dietary mineral availabilities in fish meal had any measurable effect in fish feeds. In the first study with rainbow trout, 11 supplements were tested: citric acid; sodium citrate; potassium chloride; sodium chloride; histamine dihydrochloride; EDTA disodium salt; sodium bicarbonate; a mixture of amino acids; ascorbic acid; a mixture of inositol and choline; and cholecalciferol. Apparent availability of calcium, phosphorus, magnesium, sodium, iron, manganese and strontium in fish meal-based diets was determined using both yttrium oxide (Y₂O₃) and chromium oxide (Cr₂O₃) as inert dietary markers. Apparent availability was expressed as the fractional net absorption (%) of minerals from diets. After a 7-day acclimation period with test diets, fecal samples were collected for five consecutive days using passive collection systems. Apparent availability of calcium, phosphorus, magnesium, iron, manganese and strontium was increased by citric acid supplementation. Apparent availability of manganese also was increased by EDTA and sodium citrate. The other supplements had no measurable effect on the apparent availability of minerals in fish meal. In the second study, the effect of supplemental citric acid was further investigated using monogastric (rainbow trout) and agastric fish (goldfish). Fish were fed for 5 weeks (rainbow trout) or 3 weeks (goldfish) with fish meal-based diets containing either 0% (control), 2% or 5% citric acid on a dry basis. Feces were collected by settling and by stripping. Apparent availabilities of calcium and phosphorus were greatly affected by citric acid supplementation in rainbow trout but not in goldfish. Phosphorus levels in feces of fish fed a diet with 5% citric acid were approximately half of that of fish fed the control diet (0% citric acid) in the rainbow trout trial. This pattern was consistent during the 5-week feeding trial. A dietary supplement of citric acid as high as 5% did not reduce feed intake or appetite of rainbow trout. Conversely, this level of dietary acidification led to a marked reduction of feed intake in goldfish. Dietary supplementation of citric acid at 2% level did not reduce feed intake of goldfish; however, this level of dietary acidification had little effect on the apparent availability of major minerals in fish meal-based diet. Levels of non-fecal excretion of calcium and phosphorus, inorganic phosphorus in urine, and citric acid in feces were increased in rainbow trout fed 5% citric acid. The pH values of the feces and urine were decreased in rainbow trout fed citric acid. Plasma bicarbonate, plasma calcium and phosphorus, and blood pH of rainbow trout tended to increase by a 5% dietary supplementation of citric acid. The soluble inorganic phosphorus content increased in the diets and decreased in the feces of rainbow trout by supplementing the diet with 5% citric acid. Feces samples of rainbow trout collected by stripping provided similar availability values to data collected by settling for most elements except sodium, which had negative values in all dietary treatments.     

Development and physiology of gastric dilation air sacculitis in Chinook salmon, Oncorhynchus tshawytscha (Walbaum)

The syndrome known as gastric dilation air sacculitis (GDAS) has previously been shown to affect Chinook salmon, Oncorhynchus tshawytscha, in seawater (SW) aquaculture. Feed and osmoregulatory stress have been implicated as potential epidemiological co-factors. The development and physiology of GDAS was investigated in SW and freshwater (FW) adapted smolts. Diet A (low-cohesion pellets) and diet B (high-cohesion pellets) were fed to both FW- and SW-adapted fish. GDAS was induced only in the SW trial on feeding diet A. Stimulated gastro-intestinal (GI) smooth muscle contractility, and fluid transport by the pyloric caeca were different in GDAS-affected fish, which also showed osmoregulatory dysfunction. Cardiac stomach (CS) smooth muscle contractility in response to acetylcholine and potassium chloride (KCl) was significantly reduced in fish fed diet A relative to controls from weeks 3-5. In contrast, maximal pyloric sphincter (PS) circular smooth muscle contraction in response to KCl was significantly elevated in fish fed diet A in weeks 4 and 5. Serum osmolality was elevated in GDAS-affected fish from week 2 of the SW trial. Fluid transport from the mucosal to serosal surface of isolated pyloric caeca was significantly reduced in weeks 3, 4 and 5 in SW fish fed diet A. Gastric evacuation from the stomach of healthy fish was shown to be significantly different when diets of low- and high-cohesion were fed. The results are consistent with the intestinal brake playing a role in the development of the disease.     

Effects of acclimation temperatures of 7 and 14 °C on the contractility of Atlantic salmon (Salmo salar) intestinal smooth muscle in vitro

The contractility of intestinal smooth muscle of Atlantic salmon (Salmo salar) acclimatized to temperatures of 7 and 14 °C was compared over a prolonged period of time (9 months) in freshwater. Initial studies were carried out on intestine isolated from Atlantic salmon to estimate optimal conditions for contractility. Subsequent studies were carried out with the neurotransmitter/agonist 5-HT and TS, which activates neuronal elements in the intestine, in a Krebs-Henseleit solution containing HEPES buffer maintained at pH 7.85 and at the temperature of the experimental group (i.e., 7 or 14 °C). There was a significant interaction with time and temperature for the maximal response of 5-HT (p=0.005), effective concentration producing 50% of the maximal response (EC₅₀) to 5-HT (p=0.026) and maximal response to TS (p=0.002), demonstrating that gastrointestinal contractility of Atlantic salmon is altered by both time (month) and temperature. No significant changes were found with the effective frequency producing 50% of the maximal response (EF₅₀) to TS. The potency (EC₅₀ of 5-HT) increase between 7 and 14 °C was statistically significant (p=0.026) but the small differences found with potency were most likely not physiologically significant. A time shift was recognized in the maximal responses to 5-HT, by applying a time variable to equations for the curves of the responses. This demonstrated that the curves are similar for both temperatures but alterations in the time of the maximal response changes was different. Thus, there is a delay in the time (month) changes of gastrointestinal contractility related to acclimation temperature and would be an area for future study.     

Distribution of adipocyte-related cells in skeletal muscle of rainbow trout Oncorhynchus mykiss

Adiponectin is one of the adipokines secreted mainly from adipocytes in mammals. In rainbow trout, however, adiponectin is highly expressed in skeletal muscle. Although it has been suggested that fish skeletal muscle contains adipocytes, their endocrine function and distribution are poorly understood. Recently, an EST analysis of rainbow trout found that heart-type fatty acid binding protein (H-FABP), a member of the intracellular fatty acid binding protein family, encodes an adipose-specific gene. In this study, we produced anti-adiponectin and H-FABP antibodies and investigated the distribution of adipocytes and related cells in skeletal muscle of rainbow trout. The adiponectin signal was detected at around 75 kDa in muscle in Western blotting. Since the molecular mass of rainbow trout adiponectin is around 25 kDa, this 75 kDa band would be a trimer. For H-FABP, the signal band was detected at around 15 kDa. Immunohistochemistry of skeletal muscle sections indicated that adiponectin and H-FABP signals were present outside of muscle cells and throughout the muscle tissues, suggesting the existence of adipocyte-related cells in these regions. These results will contribute to our understanding of energy metabolism in fish skeletal muscle.     

Distribution of 5-hydroxytryptamine and related compounds in various brain regions of rainbow trout (Oncorhynchus mykiss)

The levels of tryptophan (Try), 5-hydroxytryptamine (serotonin, 5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) were determined in the brain regions of rainbow trout (Oncorhynchus mykiss) by high performance liquid chromatography with electrochemical detection (HPLC-EC). Brain tryptophan concentrations varied from 3.972 ± 357 ng/g cerebellum) to 8.841 ± 772 ng/g (hypothalamus). The 5-HT concentrations varied from 69 ± 7 ng/g (optic tectum) to 573 ± 34 ng/g (hypothalamus). The concentrations of 5-HIAA varied from 29 ± 3 ng/g (medulla oblongata) to 68 ± 7 ng/g (hypothalamus). Total and free serum tryptophan levels were also determined; in adult rainbow trout 92% of the serum tryptophan was observed to be free i.e., not protein-bound.     

Semen biology of vendace (<Emphasis Type="Italic">Coregonus albula</Emphasis> L.)

The objective of this study was to describe the morphometry and motility parameters of vendace (Coregonus albula) spermatozoa. Morphometric parameters of vendace sperm head and tail were of values similar to rainbow trout. The effects of pH, sodium, potassium and calcium ion concentrations on computer-assisted sperm analysis (CASA) sperm motility characteristics were tested. Vendace sperm was motile in a wide pH range of 6.0–10.5 with the optimum pH established at 9.0. Increases in potassium and calcium ions caused decreases in the percentage of motile sperm. The CASA parameters and erratic sperm movement pattern of vendace spermatozoa were similar to whitefish (C. lavaretus) sperm motility, suggesting that there is a coregonid-specific sperm motility pattern.     

Relationships between dietary ascorbic acid status and deficiency,weight gain and brain neurotransmitter levels in juvenile rainbow trout,Salmo gairdneri

A study was conducted to determine the interrelationships between dietary ascorbic acid (AA) concentrations, brain neurotransmitter levels and weight gain in juvenile rainbow trout. At the end of 4 weeks and until the end of 12 weeks of feeding test diets of varying AA concentrations (0–320 mg AA/kg diet), increased weight gain was noted in fish fed the AA-free diet. However, by the end of 13 weeks and until the end of the experiment this phenomenon was no longer evident; instead the fish showed the more classical deficiency signs of anorexia and decreased weight gain. After 12 and 24 weeks, there were no significant differences in brain serotonin (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), norepinephrine (NE) or dopamine (DA) between fish reared on the different test diets. However, after 12 weeks of feeding the test diets, brain 5-HT, brain AA and weight gain were significantly correlated with one another. No such relationships were found for brain NE or brain DA. After 24 weeks of feeding the diets, the relationships between brain 5-HT, brain AA and weight gain were no longer apparent. Similarly, after 24 weeks brain NE and DA were also unrelated to brain AA and weight gain. These results provide evidence that in very young rainbow trout, AA deficiency, brain 5-HT levels and weight gain were related. However in fish reared on the diets for 24 weeks these relationships were no longer evident.