Cellular components of probiotics control Yersinia ruckeri infection in rainbow trout, Oncorhynchus mykiss (Walbaum)

Subcellular components of the probiotics Aeromonas sobria GC2 and Bacillus subtilis JB-1, when administered to rainbow trout, Oncorhynchus mykiss , conferred protection against a new biogroup of Yersinia ruckeri . Thus, intraperitoneal or intramuscular injection of rainbow trout with cell wall proteins (CWPs), outer membrane proteins (OMPs), lipopolysaccharides (LPS), whole cell proteins (WCPs) and live cells followed by challenge on day 8 with Y. ruckeri led to 80–100% survival compared with 10% survival in the controls. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) profiles of WCPs and OMPs from GC2 had 10 and 5 variable protein bands in comparison to 11 and 5 bands in the WCPs and CWPs from JB-1. Proteomic analyses were employed following SDS-PAGE to categorize one dominant protein of 104.7 kDa from the CWPs of JB-1 and equated it with ' Bacillus spp. endoglucanase' with a Mascot score >69. These results point to the potential of using cellular components of probiotics for protection of fish against bacterial diseases.     

Use of probiotics to control furunculosis in rainbow trout, Oncorhynchus mykiss (Walbaum)

Aerobic heterotrophic bacteria were isolated from the intestinal contents of Atlantic salmon, Salmo salar , rainbow trout, Oncorhynchus mykiss , and turbot, Scophthalmus maximus , on tryptone soya agar and De Man Rogosa and Sharpe agar, of which 11 of 177 (6% of the total) of the isolates were antagonistic to Aeromonas salmonicida . Four of these cultures, which were identified tentatively as A. hydrophila , Vibrio fluvialis , Carnobacterium sp. and an unidentified Gram-positive coccus, were beneficial to fish when fed singly or as an equi-mixture. Feed supplemented with the putative probiotics indicated survival of the organisms in the gastrointestinal tract for 7 days. Feeding with the probiotics for 7 and 14 days led to better survival following challenge with A. salmonicida . There was no indication of serum or mucus antibodies to A. salmonicida , but there was an increased number of erythrocytes, macrophages, lymphocytes and leucocytes, and enhanced lysozyme activity in the fish.     

The effectiveness of adenoviral vectors to deliver and express genes in rainbow trout,Oncorhynchus mykiss (Walbaum)

The efficacy of adenoviral vectors for gene delivery into fish cells, both in vitro and in vivo, was evaluated. Vectors utilized were of human adenovirus serotype 5 (Ad), which are commonly used in human clinical trials, but have not been assessed for gene delivery to fish. Because nothing is known about Ad receptors in fish, both an Ad (Ad5Luc1) with natural tropism for the coxsackie and adenovirus receptor (CAR), as well as an infectivity enhanced Ad (Ad5LucRGD) were included within this study. Gene expression was detected in cell lines using either vector. The levels seen with Ad5LucRGD were much higher than for Ad5Luc1 in most lines except CHSE-214. Transduction of CHSE-214 cells with Ad5Luc1 could be blocked with an excess of a competitive inhibitor, suggesting that these cells possess a CAR homologue thatmediates attachment of Ad, similar to that seen in mammalian cells. In vivo gene delivery was attempted by several methods, with significant expression seen only via intramuscular injection, although infection efficiency was low. Thus it was observed that several teleost cell lines are capable of being infected and one cell line expressed a human serotype adenoviral receptor homologue that aids in Ad infection. Additionally, in vivo studies indicated that muscle tissue of rainbow trout could be infected with Ad vectors, suggesting an alternative gene delivery strategy for this animal.     

Relationships between growth and disease resistance in rainbow trout,Oncorhynchus mykiss (Walbaum)

Rainbow trout from 23 families were evaluated for growth and resistance to the bacterial coldwater disease (BCWD) caused by Flavobacterium psychrophilum and infectious haematopoietic necrosis (IHN) caused by IHN virus. Average family weights were between 161 and 263 g with an average of 225 g at 213 days post‐fertilization with specific growth rates ranging from 2.37 to 2.88. Per cent survival of fish challenged with F. psychrophilum was between 18% and 100%, while for those challenged with IHNV, the range was between 12% and 93%. Significant positive correlations were found for end body weight and resistance to IHN (P < 0.05) and for early body weight and resistance to BCWD (P < 0.1). However, no significant correlations were detected between resistance to both pathogens or disease resistance and overall genetic diversity or diversity within the major histocompatibility locus.     

Genetic variation measured by microsatellites among three strains of domesticated rainbow trout (Oncorhynchus mykiss, Walbaum)

Genetic variation fuels selective change in natural and captive populations. In establishing a broodstock for selective improvement, the level of genetic diversity is an important consideration because it provides an indication of the scope for selective progress. Three domesticated strains of rainbow trout, Oncorhynchus mykiss (Walbaum), were examined at nine polymorphic microsatellite loci to assess detectable levels of allelic diversity and heterozygosity within and differentiation among the strains. A total of 126 alleles were observed to segregate into unique multilocus genotypes for each of the 152 individuals assayed. There was an average heterozygosity of 71.5% at these nine loci, and an average of 14 alleles at a locus. Each locus was represented by alleles unique to at least two of the three strains. Deviations from Hardy–Weinberg expectations of genotype frequencies were detected in each strain. Subsequent analysis indicated sub‐structuring within strains leading to Wahlund effects that caused these deviations. Significant differences in genotype frequencies and pairwise F_ST values demonstrated that all strains were unique. The overall F_ST of 0.089 provides additional evidence of unique genetic diversity present in each strain, and agrees well with the degree of genetic variation found in rainbow trout across broad geographical ranges. The genetic diversity contributed by each population suggests that there is greater scope for selective improvement of numerous traits within a synthetic strain combining these three strains than within any individual strain.     

Dietary modulation of digestive enzymes by the administration of feed additives to rainbow trout,Oncorhynchus mykiss Walbaum

The aim of this study was to assess the metabolic and digestive enzyme activities in rainbow trout, Oncorhynchus mykiss Walbaum fed dietary supplements. The biometric indices and the following enzymes activities were measured: acid protease and pepsin from the stomach homogenates, alkaline phosphatase (AP) from the small intestine and the brush border membrane, total proteolytic enzymes and trypsin from the small intestine and liver/pancreas. Dietary garlic induced a higher pepsin activity in the stomach than lipopolysaccharide (LPS) and ginger. Conversely, dietary ginger and LPS showed a significant difference (P > 0.05) in acid protease activity compared to the controls. There was not any significant difference observed between treated groups and the controls for hepato‐pancreas proteolytic enzyme activity, except for trypsin. AP activity increased significantly with dietary garlic, ginger and LPS compared to the controls.     

Antimicrobial peptide CAP18 and its effect on Yersinia ruckeri infections in rainbow trout Oncorhynchus mykiss (Walbaum): comparing administration by injection and oral routes

The antimicrobial peptide CAP18 has been demonstrated to have a strong in vitro bactericidal effect on Yersinia ruckeri, but its activity in vivo has not been described. In this work, we investigated whether CAP18 protects rainbow trout Oncorhynchus mykiss (Walbaum) against enteric red mouth disease caused by this pathogen either following i.p. injection or by oral administration (in feed). It was found that injection of CAP18 into juvenile rainbow trout before exposure to Y. ruckeri was associated with lowered mortality compared to non‐medicated fish although it was less effective than the conventional antibiotic oxolinic acid. Oral administration of CAP18 to trout did not prevent infection. The proteolytic effect of secretions on the peptide CAP18 in the fish gastrointestinal tract is suggested to account for the inferior effect of oral administration.     

A test of a feed budget model for rainbow trout, Oncorhynchus mykiss (Walbaum)

A feed budget model, which enables farmers to adapt a general model to local conditions, was tested on rainbow trout, Oncorhynchus mykiss (Walbaum). The model has two major components; component 1 is used to calculate the energy required for one unit of biomass gain (g or kg) and component 2 estimates the daily growth increment (g or kg). Both components can be adjusted in relation to prevailing environmental conditions and, together, they express the daily energetic requirement of the specific species or strain. Fish fed using the model as a guide had a high rate of growth and good feed conversion. The model accurately predicted growth over the experiment, and is presented as a means of calculating the daily feed budget in fish farming. Growth of fish fed using self-feeders is presented for comparison with the results obtained using the model.     

Effect of combination of dietary Bacillus subtilis and trans‐cinnamic acid on innate immune responses and resistance of rainbow trout,Oncorhynchus mykiss to Yersinia ruckeri

The present study investigated the effects of combination of dietary Bacillus subtilis and trans‐cinnamic acid on serum biochemical parameters, innate immune responses and resistance of rainbow trout, Oncorhynchus mykiss to Yersinia ruckeri. Six experimental groups of fish with mean weights of 20.58 ± 0.35 g were used in the study. Five experimental groups of fish were fed diets containing Bacillus subtilis (10⁷ per gram) or a mix of the Bacillus subtilis (BS) and trans‐cinnamic acid (25 mg/kg‐25trcBS, 50 mg/kg‐50trcBS, 75 mg/kg‐75 trcBS, 150 mg/kg‐150 trcBS), whereas an additive‐free basal diet served as the control (Cont). In this study, an increase was observed in granulocyte percentage, respiratory burst activity, phagocytic activity, phagocytic index, myeloperoxidase activity and total antiprotease activity especially in fish fed with mix of the BS and trans‐cinnamic acid‐supplemented diets (p < .05). Moreover, at the end of the 20‐day challenge period the survival rates and antibody titre (p < .05), and relative per cent survival were higher in the BS group and all trcBS groups compared with control group. As a conclusion, the results in the present study show that feeding rainbow trout with diets containing a mix of B. subtilis and trans‐cinnamic acid over a 60‐day period might be sufficient for improving fish immune responses and disease resistance against Y. ruckeri.     

Covariation between diploid and triploid progenies from common breeders in rainbow trout, Oncorhynchus mykiss (Walbaum)

The covariation between diploid and triploid progenies from common breeders was investigated in various progeny-testing experiments where either dams or sires were sampled from rainbow trout, Oncorhynchus mykiss (Walbaum), stocks. Triploidiza tion was found to frequently reduce the performance in the traits studied: body length and weight, growth, coefficient of condition and pyloric caeca number. Triploidization also generated significant interactions with the parental breeding value. These interactions were caused in part by the familial variance not being the same in triploids as in diploids, but also by actual ranking differences between diploid and triploid familial performances. However, the effect of these interactions was minor as compared with the amount of variation common to both ploidy levels (genetic correlations averaged 0.7–0.9). Therefore, selection of diploid breeders appeared efficient enough for improving triploid progeny, unless family selection methods including triploid progeny testing were preferred for other reasons. Lastly, it was observed that variances from maternal origin tended to be larger in triploids, whereas variances from paternal origin tended to be smaller, as compared with diploids. This point was discussed referring to the genetic make-up of triploids and in the scope of dams and sires for selective breeding.     

Identification of a localized mucosal immune response in rainbow trout, Oncorhynchus mykiss (Walbaum), following immunization with a protein-hapten antigen

The ability of rainbow trout, Oncorhynchus mykiss (RBT), to produce a localized mucosal immune response was investigated following intraperitoneal (i.p.) or peranal (p.a.) immunization with a protein-hapten carrier, fluorescein isothiocyanate conjugated to keyhole limpet haemocyanin (FITC/KLH). Antibody levels in serum, mucus, tissue culture supernatant from blood and spleen leucocytes, and excised skin, intestine and gill tissues were determined by ELISA. Significantly, elevated antigen-specific antibodies were elicited in both serum and mucus of fish immunized i.p. Mucosal antibody responses, in general, paralleled serum responses over time. Leucocytes isolated from spleen and blood of i.p. immunized fish at week 10 produced significantly elevated antibody levels against FITC when cultured in vitro. Excised skin, intestine and gill tissues from these fish also exhibited significantly elevated antibody responses indicating localized production in the mucosa from tissue-specific B cells. A localized mucosal immune response was elicited only after i.p. and not p.a. immunization, suggesting that systemically stimulated B cells migrate to mucosal tissues where they produce antibodies locally.     

Strain differences in feed efficiency measured as residual feed intake in individually reared rainbow trout, Oncorhynchus mykiss (Walbaum)

The efficient use of feed for growth and meat production is important for all animal production industries including aquaculture. Residual feed intake (RFI) is an alternative measure of feed efficiency that has been widely used in livestock production. Residual feed intake was calculated as the difference between intake observed and intake predicted on the basis of a bioenergetics model; a low RFI indicates greater efficiency. Residual feed intake offers some advantages as a selection criterion for improving production efficiency over traditional feed efficiency statistics because it is not a ratio and it typically has a larger coefficient of variation. The RFI of individually reared rainbow trout progeny from six different genetic cross‐types was examined for genetic variation. Proximate analysis and nitrogen retention were also evaluated to determine if differences in RFI correlate to differences in body composition and nutrient retention and varied by cross‐type. Differences between cross‐types indicated a genetic component for RFI, with the most efficient fish of approximately 160 g consuming 0.99 g less and inefficient fish consuming 0.05 g more feed per day than expected. Lower RFI was associated with higher growth rates (r=?0.38, P<0.05) and greater nitrogen retention (r=?0.82 P<0.001).     

Interaction between diet and genetic aptitude for weight and growth in juvenile rainbow trout, Oncorhynchus mykiss (Walbaum)

Samples of rainbow trout, Oncorhynchus mykiss (Walbaum) alevins from 17 independent families (full‐sib‐groups) were raised from the start of feeding until the age of 18 weeks post‐hatching with three diets (C, G and S) differing in protein content (fish soluble protein concentrate: 84% in C, 54% in G and 44% in S) and carbohydrates (none in C, 30% glucose in G and 40% crude corn starch in S). Fish were fed to near satiation, and their body weight and growth were measured. Diet effect was highly significant (G< C < S), as well as the familial effect. The major part of the familial variance (80–90%) was common to the three diets. However, a minor part of the familial variance was observed to be diet dependent (family × diet interaction), and was found to result mainly from relative performances with carbohydrates (G and S diets) vs. pure protein (C diet). These results indicate that genetic improvement of growth should suffer little impairment from possible changes in future feed formulations.