The fate of noradrenaline and adrenaline in the perfused cod spleen

The time course of appearance of catecholamine metabolites was studied in the spleen of cod,Gadus morhua, during perfusion with radioactively labelled noradrenaline and adrenaline at 10°C. The t_lag for appearance of the metabolites ranged between 1.78 and 6.76 min after onset of perfusion for both amines, indicating a rapid disposition of catecholamines. Perfusion with noradrenaline resulted in mainly MOPEG, VMA and DOMA formation, while perfusion with adrenaline additionally resulted in MN formation. There was still formation of deaminated metabolites after denervation, which indicates an additional non-neuronal site of deamination. It is concluded from the study that the fate of noradrenaline and adrenaline within the cod spleen depends on their affinities for the two uptake mechanisms and an extraneuronal site of deamination of great importance cannot be excluded.     

A specific inhibitor of mammalian kallikrein,Phe-Phe-Arg-chloromethyl ketone,inhibits the production of vasoactive substances from trout plasma by kallikrein and blocks endogenous kallikrein-like activity in trout gills

The cardiovascular effects of the kallikrein-kinin system (KKS) have not been completely characterized in lower vertebrates. In the present study, a specific, irreversible kallikrein inhibitor, Phe-Phe-Arg-chloromethyl ketone (PPACMK) was used to examine: 1. the role of the KKS in blood pressure regulation in vivo; 2. the nature of the pressor substance formed by the action of kallikrein on trout plasma in vivo and in vitro; and 3. the presence of kallikrein in trout gills and kidney. Dorsal aortic cannulated rainbow trout were used for in vivo blood pressure assays and two colorimetric serine-protease assays were used to examine tissue kallikrein activity. PPACMK alone had no effect on blood pressure in vivo. Pretreatment of porcine kallikrein with PPACMK inhibited the enzyme's pressor effect in trout by 80% and significantly attenuated the synthesis of vasopressor substance(s) from heat-treated trout plasma in vitro. Approximately 30% of gill serine protease activity was inhibited by pretreatment with PPACMK; no PPACMK-sensitive kallikrein activity was observed in the kidney. Salt water adaptation did not affect kallikrein-specific activity in the gill. These results show that the salmonid KKS does not appear to be involved in the regulation of systemic blood pressure. Further, the formation of pressor substances from trout plasma in vivo and in vitro is due to kallikrein activity and the peptides generated are similar. A kallikrein, similar to the mammalian enzyme, is found in trout gills, suggesting that the salmonid KKS may be a local mediator of gill function.     

Effects of adrenaline on ionic equilibria in red blood cells of rainbow trout (Salmo gairdneri)

Effects of adrenaline on the equilibrium distributions of Na⁺ , K⁺ , H⁺ , Cl^– , and H₂O across the cell membrane of rainbow trout (Salmo gairdneri) erythrocytes were determinedin vitro, as a function of P CO₂ (1.76–7.77 torr). CO₂-carrying capacity of the blood was also examined. Plasma catecholamine concentrations inunanaesthetized, unrestrained trout were 3.1 nM adrenaline and 1.2 nM noradrenaline. Elevation of the plasma adrenaline concentrationin vitro to 4.6 × 10³ nM resulted in net gains of Na⁺ , Cl^– and H₂O by red cells, a net loss of H⁺ from red cells, and a pronounced red cell swelling. Adrenaline also reduced the CO₂-carrying capacity of trout bloodin vitro. The magnitudes of these effects increased with P_CO2 and, thus, were sensitive to blood HCO₃ ^– concentrations. The distribution of K⁺ between red cells and plasma was unaffected by adrenaline. Adrenergic-mediated ion movements and red cell swelling were sensitive to both propranolol and SITS. These results are consistent with the symport NaCl uptake model for adrenergic-mediated swelling of Baroinet al. (1984). The adrenergic response of fish erythrocytes may function to ameliorate the effects of blood acidoses on O₂-carrying capacity by maintaining red cell pH in the face of a decrease in plasma pH.     

Anaesthetic Effects on the Hepatic Portal Vein and on the Vascular Resistance of the Tail of the Chinook Salmon (<Emphasis Type="Italic">Oncorhynchus tshawytscha</Emphasis>)

We used blood vessel myography and a perfused tail preparation from the Chinook salmon (Oncorhynchus tshawytscha) to investigate the effect of 2 commercially used fish anaesthetics, AQUI-S^TM and MS222, on vascular tone. Hepatic portal vein rings were exposed to 1 × 10⁻⁷ M adrenaline in the presence or absence of either AQUI-S^TM or MS222 and changes in vessel tension measured. Tail preparations were perfused with saline containing increasing concentrations of either anaesthetic. Exposure to either anaesthetic did not alter the response of vessel rings to adrenaline in vitro. However, both anaesthetics caused a significant (P ≤ 0.05) decrease in vessel tension when compared to controls. In tail preparations, perfusion with either anaesthetic caused a significant (P ≤ 0.05) yet reversible, dose dependent decrease in vascular resistance. Further to this, it was found that for both anaesthetics there was a significant effect of fish holding time on the response of preparations, with fish that were acclimated for less than 1 week exhibiting less vasodilation in response to anaesthetic exposure. We conclude that commonly used concentrations of AQUI-S^TM and MS222 can have a direct effect on vascular tone of salmon vessels, causing a significant but reversible vasodilation of vessels. This vasodilation may offset increases in haematocrit seen during anaesthesia in the whole animal, and in recovery prevent large rises in peripheral resistance associated with catecholamine release. A reduced vascular resistance may allow the heart to pump greater volumes of blood during recovery.     

Ammonia and urea excretion in the tidepool sculpin (Oligocottus maculosus): sites of excretion,effects of reduced salinity and mechanisms of urea transport

Tidepool sculpins live in a variable environment where water temperature, salinity, gas tensions, and pH can change considerably with the daily tide cycle. Tidepool sculpins are primarily ammoniotelic, with 8–17% of nitrogen wastes excreted as urea. The majority of net ammonia (J^net _amm; 85%) and urea (J^net _urea; 74%) excretion occurred across the gill, with the remainder excreted across the skin, the kidney, and/or gut. Acute (2h) exposure to 50% seawater significantly increased J^net _urea (2.8-fold), but reduced J^net _amm (3.5-fold). In fish exposed to 50% seawater for 1 week, J^net _urea returned to control values, but J^net _amm remained slightly depressed. Unidirectional urea influx (Jⁱⁿ _urea) and efflux (J^out _urea) were measured using¹⁴C-urea to determine if urea was excreted across the gills by simple diffusion or by a carrier-mediated mechanism. Jⁱⁿ _urea increased in a linear manner with increasing urea water levels (0–11 mmol N l^–1), while J^out _urea was independent of external urea concentrations. As well, J^net _urea and J^{out inurea} were not significantly different from one another, indicating the absence of back transport. Urea analogs and transport inhibitors added to the water did not have any consistent effect on unidirectional urea flux. These results demonstrate that ammonia and urea excretion rates and sites of excretion in tidepool sculpins are very similar to those found in other marine and freshwater teleosts. Urea and ammonia may play a role in osmoregulation as excretion rates and tissue levels were influenced by changes in water salinity. Finally, we found no evidence for a specific urea carrier; branchial urea excretion is likely dependent on simple diffusion.     

Effects of sublethal, acidic aluminum exposure on blood ions and metabolites, cardiac output, heart rate, and stroke volume of rainbow trout, Oncorhynchus mykiss

Doppler flow probes were fitted around the ventral aorta of rainbow trout, which were exposed to combinations of pH and aluminum (pH 5.1–6.2, Al 0–80 g l^–1) for 60 h. Fish accumulated Al at the gill and exhibited decreased blood Na⁺, Cl^–, and Ca²⁺ concentrations, and increased K⁺, hematocrit, hemoglobin, glucose, and lactate, indicating increasing ionoregulatory disturbance with increasing Al concentration. Fish exposed to ambient water (6.2) or low-pH (5.3) water without Al exhibited slight reductions in heart rates, as well as increased stroke volume, resulting in little variation in cardiac output. In the presence of Al (20 to 40 g l^–1) at low pH (5.1–5.3), fish increased their heart rate slightly and generally maintained their stroke volume, resulting in increased cardiac output in the first two days of exposure. At the highest Al concentration (80 g l^–1, pH 5.1), tachycardia was observed, concomitant with a decrease in stroke volume. The ionoregulatory imbalance and resulting increased blood viscosity explain these increases in heart rate rather than stroke volume in fish exposed to high concentrations of Al.     

Effects of diet and feeding time on daily variations in plasma insulin,hepatic c-AMP and other metabolites in a teleost fish,Dicentrarchus labrax L.

Juvenile sea bass, 1.5 years old, of mixed sex, held on long photoperiods were fed early on the photoperiod and late on the photoperiod, using different diets. Fish fed natural diets showed a daily rhythmicity of plasma insulin, liver c-AMP, plasma glucose, liver glycogen and muscle glycogen content, however, fish fed a commercial diet did not show this daily rhythmicity except for plasma insulin levels. In addition, these fish had significantly lower levels of plasma insulin, liver c-AMP and plasma glycerol than the group fed on the natural diets at similar feeding times. The time of feeding also induced different rhythmicity patterns in hormones and metabolites as well as a significant change in their mean levels. These facts are discussed in relation with the pre-feeding activity and increased appetite exhibited by the fish fed late on the photoperiod and with their implications on fish culture.     

Species differences in the adrenergic responses of fish red cells: studies on whitefish,pikeperch, trout and carp

The occurrence and pH dependence (pHe 7–8) of the adrenergic red cell responses of two salmonids, trout and whitefish, and a percinid, pikeperch were studied. These are all species that live in well-oxygenated waters. The responses were compared to those of carp, which tolerates oxygen-deficient waters.The adrenergic responses of trout and whitefish red cells were pronounced. In these species red cell swelling, the accumulation of sodium and chloride in the cell, and the increase in red cell oxygen content at atmospheric oxygen tension were maximal at pH 7.3. In contrast, pikeperch red cells responded to -adrenergic stimulation only at extracellular pH 7.1. In carp, the adrenergic response, occurring below extracellular pH 7.5, was small as compared to the two salmonids. In each case the onset of the adrenergic response coincided with the onset of the Root effect.The differences in the adrenergic responses between the two salmonids and pikeperch suggest that the occurrence of the adrenergic response is not directly related to the environmental oxygen requirements of the species, but may be linked to the activity pattern.     

Regulation of cardiac output and gut blood flow in the sea raven,Hemitripterus americanus

Coeliac artery blood flow (F_ca) before and after feeding was recorded in the sea raven. To obtain basic information about the scope of cardiovascular adjustment in the sea raven, a separate series of experiments was performed, in which ventral (P_va), and dorsal (P_da) aortic blood pressure, heart rate (HR) and cardiac output (jaz) were monitored during rest and encouraged exercise.Measurements of coeliac artery flow showed that visceral blood flow is substantial, particularly after feeding, and variations in the visceral vascular conductance affect P_da directly. Simultaneous recordings of intestinal and dorsal aortic blood pressures showed no measurable difference in the two arterial pressures, refuting the idea of a vascular control at the level of the main coeliac artery. Thus, in the sea raven, the adrenergic tonus affecting the visceral vasculature presumably acts at the arteriolar level.Sea ravens encouraged to exercise increased theirjaz by 64%; 32% through HR and 25% through stroke volume. The increase injaz during encouraged exercise was sufficient to produce an elevation of both P_va and P_da, despite an increase of systemic vascular conductance, -adrenoceptor blockade with sotalol, however, severely impaired the increase injaz during exercise, and the change in P_da was reversed.During rest there were both an adrenergic and a cholinergic tonus affecting the HR, as revealed by the effects of injected pharmacological antagonists. Swimming activity decreased the cholinergic tonus, while the adrenergic tonus increased.     

Involvement of cholinergic and adenosinergic systems on the branchial immune response of experimentally infected silver catfish with Streptococcus agalactiae

It has been recognized that the cholinergic and adenosinergic systems have an essential role in immune and inflammatory responses during bacterial fish pathogens, such as the enzymes acetylcholinesterase (AChE) and adenosine deaminase (ADA), which are responsible for catalysis of the anti‐inflammatory molecules acetylcholine (ACh) and adenosine (Ado) respectively. Thus, the aim of this study was to investigate the involvement of the cholinergic and adenosinergic systems on the immune response and inflammatory process in gills of experimentally infected Rhamdia quelen with Streptococcus agalactiae. Acetylcholinesterase activity decreased, while ACh levels increased in gills of infected animals compared to uninfected animals. On the other hand, a significant increase in ADA activity with a concomitant decrease in Ado levels was observed in infected animals compared to uninfected animals. Based on this evidence, we concluded that infection by S. agalactiae in silver catfish alters the cholinergic and adenosinergic systems, suggesting the involvement of AChE and ADA activities on immune and inflammatory responses, regulating the ACh and Ado levels. In summary, the downregulation of AChE activity exerts an anti‐inflammatory profile in an attempt to reduce or prevent the tissue damage, while the upregulation of ADA activity exerts a pro‐inflammatory profile, contributing to disease pathophysiology.     

The effects of fish anaesthetics (MS222, metomidate and AQUI-S) on heart ventricle, the cardiac vagus and branchial vessels from Chinook salmon (Oncorhynchus tshawytscha)

The acute effects of 3 fish anaesthetics (MS222, metomidate and AQUI-S) were investigated on 3 parts of the cardiovascular system of Chinook salmon (Oncorhynchus tshawytscha). All 3 anaesthetics reduced the contractile force of paced strips of ventricular myocardium. MS222 reduced the contractility by almost 75%, and was more potent than metomidate and AQUI-S, which reduced the contractility by about 25%. MS222 blocked vagal nerve transmission to the heart at the normally applied anaesthetic concentration (NAAC) for Chinook salmon, whereas metomidate and AQUI-S required 100 times their NAACs to have the same effect. Using myography, MS222 and AQUI-S caused a maximal 30–40% dilation of EBAs at 10% NAAC, whereas the equivalent effect with metomidate was only seen at 100% NAAC. MS222 again caused the greatest dilation of the ABAs. AQUI-S dilated the ABAs at up to 50% NAAC, but this was reversed so that there was no dilation at 100% NAAC. Metomidate did not affect the ABAs. These data from in vitro and in situ experiments, which generally show inhibitory effects, are used to suggest possible cardiovascular outcomes in anaesthetised Chinook salmon.     

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.     

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.     

The effect of acid water on oxygen consumption,circulating catecholamines and blood ionic and acid-base status in rainbow trout (Salmo gairdneri,Richardson)

Chronically cannulated rainbow trout were exposed in acid water (pH 4.0) for 72h. The gill potential was strongly dependent on water pH, being blood side negative in neutral water, but positive in acid water. Catecholamine levels increased irregularly during acid exposure, and the Bohr and Root effects were not completely erased by the effect of catecholamines during acid exposure. Long term exposure to low water pH, although causing an acidosis in the fish, did not suppress resting oxygen consumption. Prolonged exposure to acid conditions, however, resulted in an increase in ammonia excretion. Changes in plasma sodium and chloride were similar to that reported previously for trout exposed to low calcium, acid water. We conclude that exposure of trout to pH 4 soft water, although impairing oxygen transport, does not limit resting oxygen consumption but does reduce the scope for activity. More extreme acid conditions do impair resting oxygen uptake.     

The trend of lessepsian fish populations with an emphasis on temperature variations in Iskenderun Bay,the Northeastern Mediterranean

The fish assemblages of the Eastern Mediterranean are highly variable owing to the Lessepsian migration. So far, there has been limited discussion about the temporal variation of Lessepsian populations, which may be a key to better understanding the establishment, persistence and spreading dynamics of these species. The objective of this study was, therefore, to develop an understanding of inter‐annual variations of Lessepsian fish populations through investigating the effects of climate conditions. For this purpose, we evaluated the data obtained from the seasonal bottom trawl surveys conducted over the course of 12 years, from 2004 to 2015, at the infra‐littoral zone of Iskenderun Bay. Our results indicated that Lessepsians constituted 27%, 62% and 85% of total teleost fishes in the number of species (LS‐%), biomass (LW‐%) and abundance (LN‐%), respectively. The linear trends showed that the dominance of Lessepsian fishes increased with annual rates of 1.9 (p < .01, LS‐%), 2.77 (p < .01, LW‐%) and 1.43% (p < .05, LN‐%) per year during the study period. In contrast, seasonal trend decomposition based on GAMMs revealed that the true shape of trends was non‐linear. The majority of the new Lessepsians entered the study area after 2009 when warmer conditions prevailed. Those new invaders such as Nemipterus randalli, Pomadasys stridens and Apogon smithi successfully settled in a short time and remarkably altered the composition of fish assemblages after 2010. We found that variations of lessepsian fish assemblages were correlated with the increase of annual minima and average values of sea surface temperature.     

Effects of spray‐dried blood cell meal with microencapsulated methionine substituting fish meal on the growth,nutrient digestibility and amino acid retention of Litopenaeus vannamei

The effect of substitution of fish meal (FM) by spray‐dried blood cell meal (SBCM) with microencapsulated dl ‐methionine supplementation in trial diets for Litopenaeus vannamei was evaluated. Six isonitrogenous (320 g kg^?1) and isolipidic (85 g kg^?1) diets were formulated to feed shrimp (2.3±0.2 g shrimp^?1) for 56 days. Shrimp were fed with six diets in which FM protein was gradually replaced by SBCM protein (0%, 20%, 40%, 60%, 80% and 100% in diets 0–5). Growth performances and feed utilization of shrimp fed diets containing 0%, 3.5%, 7.0% and 10.5% SBCM protein were not significantly different (P>0.05). Growth, feed conversion ratio and protein efficiency ratio of shrimp fed diets (80 and 100% FM substitution) were significantly poorer compared with other treatments (P<0.05). With increased levels of dietary SBCM, apparent digestibility coefficient of dry matter, crude protein enhanced from 76.9% to 82.3%, 84.8% to 89.0%, but crude lipid decreased from 90.6% to 88.3% respectively. The carcass composition values were not significantly (P>0.05) affected by the replacement level of FM, except lipid. There were no significantly differences (P>0.05) in amino acid retentions among Diets 0–3. The results suggest that the dietary FM protein could efficiently be substituted by SBCM up to 60%, without adverse effects on the growth of L. vannamei.     

Effects of azithromycin on tilapia (Oreochromis niloticus): health status evaluation using biochemical,physiological and morphological biomarkers

Bacterial diseases cause tilapia's high‐mortality outbreak. This study investigated the toxicity of azithromycin (AZT), a macrolide antibiotic that has been considered a possible therapeutic drug for tilapia aquacultural use. The 48‐h acute toxicity (50% lethal concentration, LC50; 48 h) of AZT was determined for Oreochromis niloticus. Thereafter, fish were exposed to 0, 1, 50 and 100 mg L^?1 AZT during 14 days (chronic exposure) and measured the haematological variables, the activity of superoxide dismutase, catalase, glutathione peroxidase, glutathione‐S‐transferase (GST) and the concentration of glutathione (GSH), protein carbonyl and lipid peroxidation in the liver; histopathology was analysed the liver, gills and kidneys. The LC50; 48 h was >100 mg L^?1. No fish died during chronic exposure. Haematocrit and haemoglobin concentration increased in fish exposed to 50 and 100 mg L^?1, and the total number of leucocyte and thrombocyte increased after exposure to 100 mg L^?1 AZT, suggesting a stimulation of defence cell production. In the liver, the antioxidant enzyme activities did not change, but GST activity and the GSH level increased in fish exposed to 100 mg L^?1 AZT. Oxidative stress did not occur. Histopathological index (HI_L) indicates moderate liver damage; minor histological changes in the gill and no change in the kidneys. AZT was considered non‐toxic for O. niloticus after acute exposure and, although it causes moderated histopathology in the liver after chronic exposure, this antibiotic may be an alternative against bacterial infections, depending on its efficacy to control bacterial disease in fish.     

Effects of replacement of dietary fish oil with soybean oil on growth performance and tissue fatty acid composition in marine herbivorous teleost Siganus canaliculatus

Fish oil (FO) substitution has been studied in many marine carnivorous fish, but seldom in marine herbivorous or omnivorous species. To evaluate the feasibility of using soybean oil (SO) as a dietary lipid and confirm its capability of converting C₁₈ polyunsaturated fatty acid (PUFA) into long chain polyunsaturated fatty acid (LC‐PUFA) in the marine herbivorous teleost Siganus canaliculatus, juvenile fish were fed with four formulated diets differing in lipid composition, with SO accounting for 0.76% (SO0), 23% (SO23), 45% (SO45) and 67% (SO67) of total dietary lipid respectively. After feeding for 8 weeks, growth performance including weight gain, specific growth rate, feed conversion ratio and protein efficiency rate were better in the SO23 and, especially, SO45 groups than in the SO0 and SO67 groups (P < 0.05). Tissue fatty acid compositions were affected by diet, with the liver contents of eicosapentaenoic (EPA), docosapentaenoic (DPA), docosahexaenoic (DHA) acids and total n‐3 PUFA displaying parallel changes with the corresponding dietary fatty acids. While the muscle contents of EPA, DPA and total n‐3 PUFA between SO0 and SO23 groups, and the liver contents of arachidonic acid (ARA) and 20:4n‐3, as well as the muscle content of 20:3n‐6 between SO0 and SO45 groups showed no difference, confirming the biosynthesis of LC‐PUFA from C₁₈ precursors in vivo as the contents of corresponding fatty acids in diets SO23/SO45 were much lower than those in diet SO0 (P < 0.05). The results indicate that SO may be a suitable dietary lipid source for S. canaliculatus, and can replace up to 67% or 45% of total dietary FO without negatively compromising growth performance or nutritional quality of fish respectively. Moreover, the study increases our knowledge of FO substitution in marine herbivorous fish.