Inflammatory response of rainbow trout Oncorhynchus mykiss (Walbaum, 1792) larvae against Ichthyophthirius multifiliis

At hatching, the immune system of the rainbow trout larva is not fully developed. The larva emerges from the egg and is exposed to the aquatic freshwater environment containing pathogenic organisms. At this early stage, protection from disease causing organisms is thought to depend on innate immune mechanisms. Here, we studied the ability of young post-hatch rainbow trout larvae to respond immunologically to an infection with Ichthyophthirius multifiliis and also report on the localization of 5 different immune relevant molecules in the tissue of infected and uninfected larvae. Quantitative RT-PCR (qPCR) was used to analyze the genetic regulation of IL-1β, IL-8, IL-6, TNF-α, iNOS, SAA, cathelicidin-2, hepcidin, IL-10, IL-22, IgM and IgT. Also, a panel of 5 monoclonal antibodies was used to investigate the presence and localization of the proteins CD8, SAA, MHCII, IgM and IgT. At 10 days (84 degree days) post-hatching, larvae were infected with I. multifiliis and sampled for qPCR at 3, 6, 12, 24, 48 and 72 h post-infection (p.i.). At 72 h p.i. samples were taken for antibody staining. The first of the examined genes to be up-regulated was IL-1β. Subsequently, IL-8 and cathelicidin-2 were up-regulated and later TNF-α, hepcidin, IL-6, iNOS and SAA. Immunohistochemical staining showed presence of CD8 and MHCII in the thymus of both infected and non-infected larvae. Staining of MHCII and SAA was seen at sites of parasite localization and weak staining of SAA was seen in the liver of infected larvae. Staining of IgT was seen at site of infection in the gills which may be one of the earliest adaptive factors seen. No positive staining was seen for IgM. The study illustrates that rainbow trout larvae as young as 10 days (84 degree days) post-hatch are able to regulate important immune relevant cytokines, chemokines and acute phase proteins in response to infection with a skin parasitizing protozoan parasite.     

Effect of cellulase,phytase and pectinase supplementation on growth performance and nutrient digestibility of rainbow trout (Oncorhynchus mykiss,Walbaum 1792) fry fed diets containing canola meal

This experiment was conducted to evaluate the effects of supplementing exogenous enzymes on growth, feed conversion ratio (FCR) and apparent nutrient digestibility in rainbow trout (Oncorhynchus mykiss) fry diets containing 32% canola meal. Five experimental diets (including a control diet containing no enzymes) were prepared as isonitrogenous (44% crude protein) and isocaloric (4000 kcal DE kg¹). The four other diets contained either cellulase, phytase, pectinase or an enzyme mix (a mixture of cellulase, phytase and pectinase in the same ratio). The feeding trial was conducted in triplicate for 12 weeks in 15 tanks (100‐L). At the beginning of the experiment 20 rainbow trout fry (initial weight 1.23 g) were stocked into each tank. Mean water temperature in the rearing tanks was 11°C and water flow in each tank was 6 L min^?1. At the end of the experiment the growth parameters and FCR displayed no significant differences in enzyme supplementation (P > 0.05). In addition, no differences were observed in dry matter, protein, or lipid digestibility with enzyme supplementation (P > 0.05). The results of this study showed that the addition of pectinase, phytase, cellulase or an enzyme mix to a diet containing 32% canola meal had no effect on growth, feed efficiency or dry matter, protein, or lipid digestibility in rainbow trout fry.     

Prostaglandins in rainbow trout (Oncorhynchus mykiss Walbaum, 1792) sperm biology – searching for answers

The purpose of this study was to determine the concentrations of prostaglandins E₂ and F_2α (PGE₂ and PGF_2α) in the blood, testis and seminal plasma of mature male rainbow trout and in the ovarian fluid to assess the effects of these prostaglandins on sperm motility parameters when present in activation media. Also prolonged incubation with prostaglandins on sperm motility and calcium influx were studied. The profile of PGE₂ and PGF_2α differed in concentration between blood, testicular supernatant and seminal plasma. PGE₂ was predominant in the blood sample (0.29 ng ml^?1) and testicular supernatant (3.1 ng ml^?1) whereas their level in seminal plasma was lower than PGF_2α (0.23 ng ml^?1). The concentrations of PGF_2α in blood, testis and seminal plasma were 0.04, 0.99, 1.3 ng ml^?1, respectively. In the ovarian fluid the concentrations of both prostaglandins were higher than in the male reproductive tract. Adding both prostaglandins to activation buffer (at concentrations 15 and 70 ng ml^?1) had no effect on any CASA parameters. Calcium influx related to rainbow trout sperm incubations with PGE₂, and PGF_2α was not detected. After 24 h incubation of sperm in artificial seminal plasma solution without and with prostaglandins all sperm samples increased their motility potential and intracellular calcium concentration. Therefore, this effect was not related to the presence of prostaglandins. In summary PGE₂, and PGF_2α were present in the rainbow trout male reproductive tract, and their profile varies from that of blood, testis and seminal plasma. The specific role of both prostaglandins in salmonid sperm biology remains unclear.     

Evaluation of safflower meal as a protein source in diets of rainbow trout [Oncorhynchus mykiss,Walbaum, 1792)]*

The effects of different levels of safflower meal (a residue of the plant oil extraction process) in diets of rainbow trout (Oncorhynchus mykiss) on growth performance, nutrient digestibility and body composition were investigated. Four different diets containing 0% (Control group), 10% (Diet 1), 15% (Diet 2) and 20% (Diet 3) safflower meal were formulated for the fish with a mean weight of 132.70 ± 0.75 g fed for 70 days. At the end of the experiment, there were no significant differences among dietary treatments for weight gain, specific growth rate and feed conversion ratio (P > 0.05). There were also no differences in the digestibility of crude protein and crude lipid between groups (P > 0.05). Supplementation of safflower meal in the diets also had no adverse effects on body composition. The results indicate that safflower meal is a promising feed ingredient and can be used up to a concentration of 20% in the rainbow trout diet with no adverse effects on growth performance, nutrient digestibility or body composition.     

Hatchery performance in a major rainbow trout (Oncorhynchus mykiss,Walbaum, 1792) seed production area of Turkey

This study aims to determine the production characteristics and performance of rainbow trout hatcheries in Seydikemer, Mu?la, Turkey where over half of the total production of eyed eggs and juveniles take place. For this purpose six hatcheries with different production scales were selected and coded A, B, C, D, E and F (with capacities of 60, 15, 5.6, 4, 0.95 and 0.6 million juveniles per year, respectively). From each hatchery, 20 females were selected and their egg qualities (egg number, diameter and weight, fertilization, eyed egg, hatching, swim‐up fry and survival rates at 120 days after hatching) and juvenile growth rates were monitored until the 120^th day after hatching under each farms own conditions. The sperm characteristics of 10 broodstock males from each hatchery were also examined. Although fertilization and eyed egg rates were similar among the farms, hatching, swim‐up fry, and survival rates at the 120^th day after hatching differed significantly. The average eyed egg, hatching and survival rates (calculated from selected 20 females) were 72, 55, and 32%, respectively. While sperm characteristics except duration of motility (s), were significantly different among the farms, average sperm concentration (per ml), motility (%) and duration of motility (s) in the region were 11.8 × 10⁹, 55.3, and 56.4, respectively. The best hatchery performance was observed in Farm A because of better records and management applications than in the other farms.     

Effects of diets containing gossypol on reproductive capacity of rainbow trout (Oncorhynchus mykiss)

We evaluated five practical diets in which 0%, 25%, 50%, 75%, and 100% (dietary treatments 1-5) of fish meal protein was replaced by solvent-extracted cottonseed meal protein. Adult rainbow trout (initial average weight 247 +/- 8 g) were fed the diets over a period of 131 days during which a general 2-fold body weight increase occurred. The total diet gossypol concentration (free and protein-bound) showed a gradual increase with increased cottonseed meal substitution. Blood samples were collected on Days 0, 64, 112, and 131 for hematological and steroid hormone determination in plasma of males and females. Hemoglobin content was significantly reduced in fish from treatment 5 (7.9 +/- 0.3 g/dl) in comparison to treatments 1-3 (10.3-10.9 g/dl). After 112 and 131 days of feeding, testis weights, concentrations of testosterone, and 11-ketotestosterone were elevated in fish from dietary treatments 2 and 3 in comparison to control and diets 4 and 5. On Day 71, sperm were collected from 6 fish per dietary treatment to assess sperm quality. No significant differences in sperm concentrations (7.2-9.8 x 10(9)/ml), motility (78-89%), and standardized (300 x 10(5) sperm/egg) fertilizing ability (18.9-22.6% hatched embryos) were found. Total gossypol concentrations in blood plasma differed significantly among treatments, and the levels were among the highest ever recorded in animals fed cottonseed-supplemented diets (2.9 +/- 0.2, 11.7 +/- 4.1, 21.7 +/- 1.4, and 29.9 +/- 3.9 microg/ml, for treatments 2-5, respectively). The major portion of gossypol in blood plasma was protein-bound (81-93%). This was in contrast to minute amounts of gossypol present in seminal plasma, mostly in free form (0.02-0.18 microg/ml), which indicates the presence of a barrier between general circulation and the testis with respect to gossypol distribution in lower vertebrates. Thus, the reproductive parameters of male rainbow trout examined in this study were not significantly affected by feeding cottonseed meal for 131 days.     

Environmental influences on cardiovascular variables in rainbow trout, Oncorhynchus mykiss (Walbaum)

Two groups of wild (lakedwelling and anadromous), and a group of hatchery-reared Oncorhynchus mykiss (Walbaum) (rainbow trout) were sampled in order to measure cardiac morphometrics, haemoglobin concentration, and the DNA and protein concentration in cardiac muscle. A combination of these variables was used to distinguish wild fish from domestic ones.
The wild fish had significantly higher levels of haemoglobin (for male fish, 10.10 and 10.07 g 100 ml⁻¹ vs. 7.69 g d⁻¹) and larger relative ventricle mass (females, 0.091 and 0.099% ofbody mass vs. 0.073%; males, 0.108 and 0.134% vs. 0.102%; immatures, 0.086 and 0.094% vs. 0, 072%, respectively) than the domestic fish. The anadromous and domestic fish had significantly higher amounts of compact tissue when compared with lake fish (females, 43 and 47% of ventricle mass vs. 34%, respectively). Ventricle size distinguished wild fish from domestic fish, except male anadromous and male domestic fish which were distinguished only by haemoglobin and compact tissue values. Immature fishes from all groups had lower total protein levels in the ventricle, lower compact tissue levels, and less haemoglobin. Points regarding the potential environmental influences in determining these cardiovascular trends are discussed.     

Haemoglobin isomorph abundances in splenectomized rainbow trout, Oncorhynchus mykiss (Walbaum)

Rainbow trout, Oncorhynchus mykiss (Walbaum) were rendered acutely anaemic by immersion in phenylhydrazine HC1 at 5° C. Following splenectomy or sham operation erythropoiesis was induced by gradual warming. No consistent or significant differences in red cell numbers, haemoglobin, haematocrit, mean erythrocytic volume or haemoglobin or haemoglobin isomorph system organization were evident during or following recovery. It is unlikely, therefore, that reported acclimatory adjustments in haemoglobin isomorph abundances can be attributed to site-related differences in erythroid cell lineages.     

The effects of different cycles of starvation and re‐feeding on growth and body composition in rainbow trout (Oncorhynchus mykiss,Walbaum, 1792)

This study was designed to investigate the effects of starvation and re‐feeding cycles on the growth performance and body chemical composition of Oncorhynchus mykiss juveniles. A total of 360 juveniles with initial mean weights (IW) of 8.46 ± 0.07 g (n = 360) were stocked into 400‐L tanks in triplicate for each group, with 30 juveniles per tanks. The control group received regular feed, as is the common practice. The three other groups were periodically starved: 1 day starvation followed by 6 days re‐feeding (S1), 2 days starvation followed by 5 days re‐feeding (S2) and 3 days starvation followed by 4 days re‐feeding (S3). The experiment lasted for 10 weeks, over the course of which the water flow rate was 4 L min^?1 and the water quality parameters determined as: temperature 14.4 ± 1.1°C, oxygen 8.2 ± 0.4 mg L^?1 and pH 7.5 ± 0.2. At the end of the study, S1 had the best growth performance (final weight, specific growth rate, average daily growth) of all test groups (P < 0.05). The lowest daily feed intake (DFI) and growth performance parameters were observed in S3 (P < 0.05), while protein efficiency ratio (PER), net protein utilization (NPU) and lipid efficiency ratio (LER) were higher in the S3 fish group than in the other groups (P < 0.05). Whole body protein and lipid contents were highest in S1 fish. The hepatosomatic index (HSI) and viscerosomatic index (VSI) were significantly different among groups (P < 0.05). Feed conversion ratio (FCR) was significantly lower in starvation groups S1, S2 and S3 than in the control (P < 0.05). Compensation coefficient (CC) values were higher than 1 in all starvation groups. The concluding indicate that rainbow trout exposed to 1 and 2 days of starvation in week cycles could achieve over compensation compared to the control. Additionally, partial growth compensation and improved feed utilization could be achieved in a starvation group within 3 days in a week, by beginning with the juvenile size over a 10‐week experimental period.     

The effect of mycotoxin deoxynivalenol (DON) on the oxidative stress markers in rainbow trout (Oncorhynchus mykiss,Walbaum 1792)

Deoxynivalenol (DON) is one of the most frequently detected mycotoxins in agricultural commodities used as animal feedstuff in Central Europe. This study focuses on determining effects of diets containing DON on oxidative stress markers and detoxifying enzymes in rainbow trout (Oncorhynchus mykiss). The fish were fed with commercial pellets containing DON at a concentration of 2 mg kg^?1. Selected enzymes were measured in liver, gill and caudal kidney of the fish after 23 and 32 days of the experiment. Significant differences between the control and experimental groups were observed concerning activities of glutathione peroxidase (GPx) in kidney, glutathione reductase (GR) in gill and kidney, catalase (CAT) in kidney and liver and glutathione S‐transferase (GST) in gill and liver. No significant differences were found for superoxide dismutase (SOD) gene expression, lipid peroxidation (TBARS) and the ferric reducing ability of plasma (FRAP). The data show that DON in the diet at the concentration below EC recommendation (2006/576/EC) induces oxidative stress in the rainbow trout.     

Opsonizing properties of natural antibodies of rainbow trout, Oncorhynchus mykiss (Walbaum)

In order to confirm previous observations in which a protective effect of rainbow trout natural antibodies against furunculosis was suspected, phagocytosis studies wereconducted in vitro , using combinations of rainbow trout sera with high or low levels of natural antibodies and active or inactivated complement as opsonizing factors. Opsonization was observed in all the cases where complement was present, and to a lesser degree with sera containing only natural antibodies. The results confirm the prime importance of the complement system and provide additional evidence for a possible role of natural antibodies in antimicrobial defences.     

The ontogeny of drinking in the rainbow trout, Oncorhynchus mykiss (Walbaum)

Drinking was measured by the rate of uptake of tritiated dextran in fresh water by the alevins and fry of rainbow trout at various stages of development during a 40-day post-hatch period. Drinking increased almost S-fold during the initial 16 day yolk-sac stage. Drinking rate increased most between 16 and 23 days, the transitional period between yolk-sac absorption and first feeding. The maximum weight-specific drinkingrateof 3.24μlg⁻¹ h⁻¹ recorded for 40-day-old fry was higher than previously recorded for adults. Abrupt transfer from an adaptation temperature of 10 to 19° C increased drinking significantly (Q₁₀= 1.2), but sudden transfer to 1 1‰ salinity sea water caused a substantial fall in drinking rate in 23-day fry. A 24-h period of adaptation to 1 1‰ and 15‰ salinity restored drinking to a rate similar to that in fresh water. The water drunk by 31-day fry fed to satiation was initially higher than by unfed fry, but drinking rates subsequently fell below the control level. The results are discussed in terms of osmoregulation and the uptake of dissolved or suspended substances by the intestinal tract for the purpose of immunization.     

Kocuria SM1 controls vibriosis in rainbow trout (Oncorhynchus mykiss,Walbaum)

Aims: To develop probiotics for the control of vibriosis caused by Vibrio anguillarum and Vibrio ordalii in finfish. Methods and Results: Kocuria SM1, isolated from the digestive tract of rainbow trout, was administered orally to rainbow trout (Oncorhynchus mykiss) for 2 weeks at a dose equivalent to c. 10⁸ cells per g of feed and then challenged intraperitoneally with V. anguillarum and V. ordalii. Use of SM1 led to a reduction in mortalities to 15–20% compared to 74–80% mortalities in the controls. SM1 stimulated both cellular and humoral immune responses in rainbow trout, by elevation of leucocytes (5·5 ± 0·8 × 10⁶ ml^?1 from 3·7 ± 0·8 × 10⁶ ml^?1), erythrocytes (1·2 ± 0·1 × 10⁸ ml^?1 from 0·8 ± 0·1 × 10⁸ ml^?1), protein (23 ± 4·4 mg ml^?1 from 16 ± 1·3 mg ml^?1), globulin (15·7 ± 0·2 mg ml^?1 from 9·9 ± 0·1 mg ml^?1) and albumin (7·3 ± 0·2 mg ml^?1 from 6·1 ± 0·1 mg ml^?1) levels, upregulation of respiratory burst (0·05 ± 0·01 from 0·02 ± 0·01), complement (56 ± 7·2 units ml^?1 from 40 ± 8·0 units ml^?1), lysozyme (920 ± 128·8 units ml^?1 from 760 ± 115·3 units ml^?1) and bacterial killing activities. Conclusions: Kocuria SM1 successfully controlled vibriosis in rainbow trout, and the mode of action reflected stimulation of the host innate immune system. Significance and Impact of the Study: Probiotics can contribute a significant role in fish disease control strategies, and their use may replace some of the inhibitory chemicals currently used in fish farms.     

Red beet and betaine as ingredients in diets of rainbow trout (Oncorhynchus mykiss): effects on growth performance,nutrient retention and flesh quality

The objective of the study was to evaluate the impact of different concentrations of dietary red beet and betaine on the growth performance and fish flesh quality of rainbow trout. Therefore, a control diet was compared with four diets in which two levels of red beet (14% and 28%) and betaine (0.9% and 1.63%) were incorporated in combination. The study was set up with an average body weight of 69 ± 2.2 g and finished when fish reached commercial weight (175–250 g) after 105 d. The impact of the diets was studied based on the growth performance, biometric indexes, proximal composition, protein and fat retention efficiencies and apparent nutrient digestibility by fish reared on a recirculation system. Further estimates were the effect of red beet and betaine on the flesh proximate composition and quality of the final product (water activity, colour, texture, thiobarbituric acid reactive substances and sensory characteristics). Results showed that inclusion of 14% red beet and 0.9% betaine did not affect growth, nutritive or biometric parameters and nutrient retention when compared with the control diet. However, higher levels of red beet and betaine had negative effects on growth and nutritive parameters. The tested ingredients enhanced quality parameters regardless of the concentration used. After feeding the red beet and betaine, fish flesh showed lower water activity and better textural and colour properties than the control and also a dose-dependent effect on lipid oxidation was observed.     

Dichloroacetate reveals the presence of metabolic inertia at the start of exercise in rainbow trout (Oncorhynchus mykiss,Walbaum 1792)

A lag in the increase in oxygen consumption (MO₂) occurs at the start of sustainable exercise in trout. Waterborne dichloroacetate (0.58 and 3.49 mmol l⁻¹), a compound which activates pyruvate dehydrogenase (PDH) by inhibiting PDH kinase in muscle, accelerates the increase in MO₂ during the first 10 min of sustainable exercise when velocity is elevated to 75% critical swimming speed in a swim tunnel. There are no effects on MO₂ thereafter or at rest. This indicates that a delay in PDH activation (“metabolic inertia”) contributes to the lag phenomenon.     

A blue variant in the rainbow trout, Oncorhynchus mykiss Walbaum

A blue variant of the rainbow trout, which appeared in a French fish farm, displayed an iridescent body color that was cobalt blue on the back, lighter on the undersides, and silvery on the belly and which held up to adult stage. This color was supposed to result from a Tyndall effect involving a structural arrangement of melanin pigments because it disappeared when it was associated with a depigmenting gene. This blue variant appeared to be governed by an autosomal recessive gene. Blue fry survival and body weight were about 25% less than those of wild-type sibs, but no major problem was observed in further breeding performances, including reproduction. These features do not correspond with those of the blue variants previously described in the rainbow trout.     

Pathogenicity of vibrios to rainbow trout (Oncorhynchus mykiss, Walbaum) and Artemia nauplii

The taxonomy of marine vibrios has changed rapidly over the last two decades, and a wealth of new species have been identified. Many Vibrio species are pathogenic to fish and crustaceans; however, little is known about the virulence of many of the novel species. The purpose of this study was to determine the ability of various recent isolates of vibrios to cause disease in rainbow trout (Oncorhynchus mykiss, Walbaum) and crustacea, i.e. Artemia nauplii. Of 56 isolates, representing 26 species of Enterovibrio, Photobacterium and Vibrio, obtained from a diversity of healthy and diseased aquatic animal hosts and water samples from many geographical locations, Vibrio brasiliensis, V. coralliilyticus, V. ezurae, V. fortis, V. kanaloaei, V. neptunius, V. rotiferianus and V. tubiashii were pathogenic to rainbow trout and Artemia nauplii with mortalities of up to 100%. The extracellular products of these pathogenic isolates were harmful to the animal models. In contrast, cultures of Enterovibrio norvegicus, E. coralii, Photobacterium rosenbergii, Vibrio campbellii, V. chagasii, V. cyclitrophicus, V. gallicus, V. gigasii, V. hepatarius, V. hispanicus, V. lentus, V. nereis, V. pacini, V. pomeroyi, V. shilonii, V. superstes, V. tasmaniensis and V. xuii demonstrated either non- or low virulence in the animal models.     

Innate immune responses in rainbow trout (Oncorhynchus mykiss, Walbaum) induced by probiotics

Carnobacterium maltaromaticum B26 and Carnobacterium divergens B33, which were isolated from the intestine of healthy rainbow trout (Oncorhynchus mykiss, Walbaum), were selected as being potentially useful as probiotics with effectiveness against Aeromonas salmonicida and Yersinia ruckeri. Thus, rainbow trout administered with feed supplemented with B26 or B33 dosed at >10(7) cells g(-1) feed conferred protection against challenge with virulent cultures of the pathogens. Moreover, both cultures persisted in the gut for up to 3 weeks after administration. The cultures enhanced the cellular and humoral immune responses. Specifically, fish fed with B26 demonstrated significantly increased phagocytic activity of the head kidney macrophages, whereas the use of B33 led to significant increases in respiratory burst and serum lysozyme activity. Also, the gut mucosal lysozyme activity for fish fed with both cultures was statistically higher than the controls.