Salmo Irideus Research Articles

Bacteriophage Cocktail Effectively Prolonging the Shelf-Life of Chilled Fish.

We developed a technological accessory bacteriophage-based preparation and a method for phage-mediated bioprocessing for elimination of pathogenic microorganisms from the surface of fresh fish and for prolongation of the shelf-life of chilled hydrobionts. Specimens of rainbow trout (Salmo irideus) served as the objects of the study carried out at a fish-processing plant in the Republic of Karelia. The specimens were decontaminated by a bacteriophage cocktail containing six original virulent phage strains characterized by their pheno- and genotypical properties. A new method of biodecontamination (plunging the rainbow trout for 30 sec into a solution of bacteriophage cocktail (bacteriophage titers ≥108 PFU/ml) delayed bacterial degradation of hydrobionts by 3 days. The use of the new method for decontamination of food half-products - phage-mediated bioprocessing - promoted preservation of the initial ecological purity, nutritive value, and taste of the products and prolonged their shelf-life in comparison with the actual standards.

Oxidative stress and band 3 protein function in Liza aurata and Salmo irideus erythrocytes: effect of different aquatic conditions

The aim of this study was to assess the effect of the different aquatic conditions on anion transport of fish erythrocytes through the measurement of the sulphate uptake operating from band 3, the determination of reduced glutathione (GSH) and oxidized glutathione (GSSG). To this purpose, blood samples of 30 Liza aurata and 30 Salmo irideus fishes were collected via caudal vein and washed with physiological buffer. Successively, erythrocytes suspended at 3% haematocrit were used to measure the SO(4) (=) influx by atomic absorption spectrophotometry at 425 nm wavelength and the GSH concentration using an immunodiagnostic assay intended for the quantitative determination of glutathione in ethylenediaminetetraacetic acid in blood. All results were analyzed using unpaired Student's t-test and a P < 0.05 was considered statistically significant. The results of this study showed statistically significant differences about the sulphate uptake and the glutathione levels in S. irideus with respect to L. aurata (P < 0.0001). In conclusion, the different aquatic conditions play an important role on anion transport in fish erythrocytes, showing that environmental differences induce perturbations in erythrocyte membrane and should be evaluated to prevent physiological damages in fishes.

Structural changes in the erythrocyte membrane of the trout Salmo irideus at seasonal acclimatization

Differences of thermostability were studied in red blood cells of the trout Salmo irideus differing in sex and age, as well as structural-dynamic characteristics of erythrocyte membrane proteins at seasonal acclimatization in the interval of reservoir water temperature of 0–19°C. An increase of resistance of erythrocytes to temperature lysis with elevation of the environmental temperature was revealed to be accompanied by a rise of the proteins segmental mobility and a decrease of intermolecular interactions in spectrin-actin cytoskeleton from the data of the ESR spin labeling method. Regulation of erythrocyte stability during acclimation was concluded to occur both changes of the fatty acids chain package at the variations of lipid composition and by changes of the cytoskeleton structural lability. Thereby this provides an increase of the bilayer firmness, on the one hand, while, on the other hand, a rise of elasticity and expansibility of the membrane on the whole, which increases resistance of cells to colloidal-osmotic hemolysis. Changes of concentration of oxygen dissolved in water, which are caused by temperature fluctuations, do not deem to be of crucial importance for structural stability of erythrocytes, as it can be compensated by another mechanism, specifically by changes of affinity of hemoglobin to oxygen.

The effect of some tannins on trout erythrocytes exposed to oxidative stress

In order to gain more knowledge on the role of tannins as antioxidants, their ability to protect ( Salmo irideus) erythrocytes against oxidative stress was investigated. Antioxidant activity of different tannins (tannic, gallic and ellagic acid) was evaluated by chemiluminescence (CL) techniques using lucigenin and luminol as chemiluminogenic probes for the superoxide radical generated by the xanthine/xanthine oxidase system and hydrogen peroxide, respectively. The superoxide-scavenging activity of these tannins was shown for all the compounds; however, it is not clear if this is due to their ability of scavenging the superoxide radical or to their inhibitory activity on xanthine oxidase. Tannic and ellagic acid showed a marked effect on the reduction of H 2O 2-luminol chemiluminescence. The influence of these tannins on the rate of hemolysis in stressed trout erythrocytes was investigated and the results indicate that tannic acid accelerates the hemolytic event while gallic and ellagic acid have no significant effect. The possible protective action of these compounds against oxidative DNA damage was assessed using the comet assay, a rapid and sensitive single-cell gel electrophoresis technique, used to detect primary DNA damage in individual cells. The results here reported show that tannins under study are capable at low concentrations of protecting DNA breakage, while at high concentrations they can be genotoxic.

Correlation between functional and structural changes of reduced and oxidized trout hemoglobins I and IV at different pHs. A circular dichroism study.

Circular dichroism (CD) spectra of two major hemoglobin components (Hb), HbI and HbIV, from Oncorhyncus mykiss (formerly Salmo irideus) trout were evaluated in the range 250-600 nm. HbI is characterized by a complete insensitivity to pH changes, while HbIV presents the Root effect. Both reduced [iron(II) or oxy] and oxidized (met) forms of the two proteins were studied at different pHs, 7.8 and 6.0, to obtain information about the pH effects on the structural features of these hemoglobins. Data obtained show that oxy and met-HbI are almost insensitive to pH decrease, remaining in the R conformational state also at low pH. On the contrary, the pH decrease induces similar structural changes, characteristics of ligand dissociation and R-->T transition, both in the reduced and in the oxidized HbIV. The structural changes, monitored by CD, are compared with the peroxidative activity of iron(II)-Hb and met-Hb forms and with the superoxide anion scavenger capacity of the proteins.

Biochemical Properties of Lipid Composition of Lipoproteins in Fish of Different Ecological Groups, Rainbow Trout Salmo irideus L. and Whitefish Coregonus lavaretus L.

We have studied the lipid composition of low- and high-density lipoproteins from the blood sera of female rainbow trout Salmo irideus L. and whitefish Coregonus lavaretus L. Phospholipids prevail in all lipoprotein fractions, and phosphatidylcholine is dominant among them. Species-specific features of lipid fractions depend on ecological conditions under which these fish exist: rainbow trout is an almost domesticated species, whereas whitefish is a component of the natural ichthyofauna. The sera of these species contains lipoprotein groups specific for other animals; however, high-density lipoproteins prevail in fish.

Hemoglobin components from trout ( Salmo irideus): determination of their peroxidative activity

The peroxidative activity of trout hemoglobins, HbI and HbIV, which differ in their conformation, was compared with that of HbA. Artificial substrates (guaiacol and dopamine) and more physiological substrates such as model lipid membranes containing unsaturated fatty acids were used. The results indicate that all the hemoglobin molecules assayed show different levels of peroxidative activity. The capability to act as peroxidases is greater in HbIV than in HbI and HbA. In contrast, native globins did not show peroxidase activity. The different peroxidative activity of the Hbs is discussed in relation to stability both vs. protein oxidation and protein dissociation. The results confirm the view that hemoglobin may be of importance in establishing the life span of the erythrocyte itself.

DNA damage induced by organotins on ­trout‐nucleated erythrocytes

AbstractThe effect of tributyltin‐chloride (TBTC), dibutyltin‐chloride (DBTC) and monobutyltin‐chloride (MBTC) on rainbow trout (Salmo irideus) nuclear DNA, was investigated by means of single cell gel electrophoresis (‘comet’ assay). Our data show that TBTC presents a marked genotoxic effect, whereas the genotoxic effect is less pronounced for DBTC and it is completely absent for MBTC. These results could be important in evaluating the environmental risks deriving from the use of these molecules as a antifouling agents in marine paints and as agricultural biocides. Copyright © 2001 John Wiley &amp; Sons, Ltd. Abbreviations: TBTC, tributyltin‐chloride; DBTC, dibutyltin‐chloride; MBTC, monobutyltin‐chloride; Hb, hemoglobin.

Thermoresistance of erythrocytes and hemoglobin during acclimation of the rainbow trout Salmo irideus

Changes of parameters of thermally induced hemolysis of erythrocytes and of thermal denaturation of hemoglobin were studied at acclimation of the rainbow trout Salmo irideus to seasonal changes of water temperature in the range of 0–20°C. A correlation was revealed between thermoresistance of erythrocytes, activation energy of thermal hemolysis, and water temperature variations, whereas thermostability of hemoglobin remained practically constant. The conclusion is made that seasonal changes of thermoresistance of erythrocytes are determined by rearrangements in membrane, which play adaptive role and are due to variations in lipid composition.

Effect of three diaryl tellurides, and an organoselenium compound in trout erythrocytes exposed to oxidative stress in vitro

Previous literature reports have demonstrated that nucleated trout erythrocytes in conditions of oxidative stress are subjected to DNA and membrane damage, and inactivation of glutathione peroxidase. The present study was undertaken to evaluate the ability of three diaryl tellurides and the organoselenium compound ebselen to protect trout ( Salmo irideus) erythrocytes against oxidative stress, induced thermally and by a variation of pH. The antioxidant ability of these molecules was evaluated through chemiluminescence. Impairment of DNA was assessed using the comet assay, a rapid and sensitive single cell gel electrophoresis technique, used to detect primary DNA damage in individual cells. At low concentrations (<10 μM), all the compounds used presented a protective effect on DNA damage without altering the hemolysis rate. In higher concentrations, they accelerated the hemolysis rate and two of the diaryl tellurides were strongly genotoxic.

Changes in Physiological Indicators and Behavioral Responses Induced by Pinealectomy in Rainbow Trout (Salmo irideus)

Changes in Physiological Indicators and Behavioral Responses Induced by Pinealectomy in Rainbow Trout (Salmo irideus)

Plasma membrane perturbation induced by tributyltin chloride on density-separated trout erythrocytes

By using a discontinuous Percoll gradient in the 45–65% range it is possible to separate nucleated Salmo irideus erythrocytes in three different density fractions that are related to aging of the cell. The stability to lysis of the erythrocytes obtained from the three layers was examined in the presence and in the absence of Bu3SnCl4 (TBTC). The rate of hemolysis in the absence of the organotin was nearly the same for the three erythrocyte fractions. The addition of TBTC to the suspension increased the hemolysis but the effect depended on the density of the fraction. The change in the hemolysis rate was in the order bottom (B) > middle (M) > top (T). The effect of TBTC on 1,6-diphenyl-1,3,5-hexatriene (DPH) steady-state fluorescence anisotropy was measured in liposomes formed by the lipids extracted from the different fractions. Compared with the controls, the DPH anisotropy decrease in the presence of TBTC. Steady-state fluorescence of 2-dimethylamino-6-laurylnaphthalene (Laurdan) was measured to evaluate membrane polarity. The presence of TBTC on the same liposomes decreased the generalized polarization (GP340) in the top and middle fractions while it did not change in the bottom fraction. Copyright © 1999 John Wiley & Sons, Ltd.

Phospholipids and their fatty acids in mitochondria, synaptosomes and myelin from the liver and brain of trout and rat: a new view on the role of fatty acids in membranes

The content of different phospholipids (PL) and their fatty acid (FA) composition in subcellular fractions from the liver and brain of rat ( Rattus rattus) and trout ( Salmo irideus) were estimated. It was shown that despite higher content of unsaturated fatty acids in myelin compared to synaptosomes, the unsaturation index of the latter is equal or higher than that of myelin. The total content of PL polyunsaturated fatty acids (PUFA) was shown to be higher in membrane structures with more active ion transport (mitochondria). This feature seemed to be characteristic of membranes from both representatives of homoiotherms and poikilotherms studied. A possible role for PUFA in formation within the lipid monolayer of areas with different capacity to accept electrons and transport them along a sort of intermolecular ‘tunnel’ is discussed. The double bonds of PUFA in this area seem to be able to produce bonds similar to conjugated bonds.

Light-dependent Changes in Physiological Indices and Behavior Activity of the Rainbow Trout Salmo irideus

Light-dependent Changes in Physiological Indices and Behavior Activity of the Rainbow Trout Salmo irideus

The Effect of Indolinic and Quinolinic Nitroxide Radicals on Trout Erythrocytes Exposed to Oxidative Stress

The purpose of this study was to evaluate the ability of indolinic and quinolinic nitroxide radicals to protect trout (Salmo irideus) erythrocytes against oxidative stress. By using laurdan as a fluorescence probe, it was observed that the nitroxides inhibited the shift towards a gel phase of liposomes prepared with phospholipids extracted from trout erythrocyte membranes prior to the hemolytic event. In addition, the presence of 100 μM nitroxides in these liposomes protected the latter against lipid peroxidation determined by monitoring conjugated diene formation. However, the short chain analogue of the indolinic nitroxide and the quinolinic nitroxide had a negative effect on trout hemolysis, contrary to what has already been observed in previous studies on human RBCs (red blood cells). The half-time (t1/2) of the hemolytic process was 174 ± 4.02 min for the former and 184 ± 4.30 min for the latter compared to the control, 283 ± 5.05 min. Furthermore, the nitroxides remarkably increased the autoxidation rate of both trout and human hemoglobin to met-Hb. Even though protection at the membrane level is conferred by the nitroxides during the early stages of lipid peroxidation, their antioxidative ability might be overwhelmed at a later stage by other mechanisms such as the increased autoxidation of hemoglobin in the presence of the nitroxides, thus giving a possible explanation for the early induction of hemolysis induced by the nitroxides. The superoxide scavenging ability of all the nitroxides used was also evaluated through chemiluminescence.

Interaction of trout hemoglobin with H2O2: a chemiluminescence study.

Erythrocytes from trout Salmo irideus are characterized by four different hemoglobin components (HbI, HbII, HbIII and HbIV), HbI and HbIV being predominant. In this study we describe the interaction between trout hemoglobin (HbI and HbIV) and H2O2 using a chemiluminescence assay. Our data show that the reaction of hemoglobins with H2O2 produces a time-limited and significant increase of chemiluminescence signal. The half-life of the decay of this chemiluminescence signal was characteristic for each type of hemoglobin used. These results indicate the formation of excited molecules related to the interaction between trout hemoglobin and H2O2.

On the Origin of Interindividual Susceptibility to Motion Sickness

According to the otolith asymmetry hypothesis for susceptibility to motion sickness, the interlabyrinthine asymmetry in the otoconial mass, which is normally compensated in the circuity of the vestibular system by appropriate neuronal weighting, becomes disrupted as a result of unfamiliar movement patterns or force environments. Measurements in various species have demonstrated a large scatter in the otoconial mass. As the utricle and saccule have distinct functions in the vestibular system it seems appropriate to deal with these organs separately. Results are presented on mass distribution and lateral preponderance of the otoconial mass in the utricles and saccules of the salmon (Salmo salar) and trout (Salmo irideus). The measurements revealed considerably larger dimensions of the saccular otoconia in these species, amounting to twenty-fold. This substantial difference indicates that different regulatory principles underlie the otoconial generation of each of the organs and is presumably related to their specific functions. The lateral preponderance was found to be normally distributed for both organs in both species, with standard deviations of approx. 4% except for the utricular otoconia of the salmon (13%). In a second set of experiments, fish were exposed to a Coriolis force environment. Their sealed aquarium was subjected to constant vertical axis rotation combined with pendular oscillation around the horizontal axis. The aquarium was illuminated by a light source fixed to project through the top, and a video camera recorded the movements of the fish. During combined rotation and pendular oscillation, one group offish maintained an active compensatory swimming behaviour, whereas the movements of a second group became uncoordinated and often led to passive behaviour. Analysis of the otoconia of these two groups of fish promises to yield further evidence relevant to the otolith asymmetry hypothesis.

Plasma Membrane Perturbation Induced by Organotins on Erythrocytes fromSalmo irideus Trout

Tributyltin chloride and its degradation products monobutyltin and dibutyltin act as water pollutants, owing to the use of tributyltin chloride as a biocide in marine paint formulations. These compounds are lipid-soluble and undergo bioaccumulation and bioconcentration. Salmo irideus trout erythrocytes were studied to evaluate the possible effects of these compounds on freshwater fish, which could be exposed to long-term effects due to bioaccumulation of organotins. Data showed that tributyltin increases the haemolysis rate, starting at 10 μM, while dibutyltin has a scant protective effect at each concentration tested. Similar studies were performed in the presence of carbon monoxide (CO), which is protective against membrane oxidative stress due to haemoglobin (Hb) auto-oxidation. In these conditions all the organotins tested induced an increase in the haemolysis rate. These results suggest that the consequence of auto-oxidation of Hb could condition the effects of some organotin compounds. Steady-state fluorescence of probes embedded in the lipidic part of the membrane was used to evaluate the modifications induced by organotins to the physico-chemical state of phospholipids.

Seasonal variations of physical and biochemical membrane properties in trout erythrocytes ( Salmo irideus)

Erythrocyte membranes from Salmo irideus trout were analysed month by month, focusing on (1) Na +,K +-ATPase activity; (2) cholesterol content; (3) generalised polarization of laurdan, reflecting membrane lipid polarity and (4) 1,6-diphenyl-1,3,5-hexatriene steady-state fluorescence anisotropy, related to “membrane fluidity.” Properties (2) and (3) showed clear maxima during winter, property (1) during summer, whereas property (4) was not much influenced by season. Clearly, Na +,K +-ATPase activity of trout erythrocytes is influenced by the physical state of membrane lipids on a monthly scale.

Determination of protein quality of rainbow trout ( Salmo irideus) by in vitro protein digestibility — corrected amino acid score (PDCAAS)

Protein digestibility and protein quality of raw rainbow trout, broiled rainbow trout and smoked rainbow trout were studied by in vitro assay, Amino acid score (AAS) and protein digestibility corrected amino acid score (PDCAAS). Protein digestibilities of samples were determined using an in vitro, three-enzyme method in a pH-stat and three- and four-enzyme pH-drop methods. Amino acid score was based on the amount of the single most limiting amino acid, and its calculation included the use of the requirement pattern suggested by FAO/WHO/ UNU for pre-school children. Protein digestibilities of raw, broiled and smoked rainbow trout were found to be 87.1, 84.0 and 83.4% using the three-enzyme pH-drop method, 84.7, 81.4 and 80.8% using the four-enzyme pH-drop method, and 95.5, 93.9 and 91.2% using the three-enzyme pH-stat method, respectively. When the amino acid score was corrected for in vitro (three-enzyme pH-stat method) protein digestibility, the resulting values of 99.8, 97.1 and 93.9% were obtained. Amino acid score corrected for protein digestibility seems to predict, accurately, the nutritional quality of fish protein when in vitro values are used.