Innovations in fish nutrition act as drivers for the sustainable development of the rapidly expanding aquaculture sector. Probiotic dietary supplements are able to improve health and nutrition of livestock, but respective bacteria have mainly been isolated from terrestrial, warm-blooded hosts, limiting an efficient application in fish. Native probiotics adapted to the gastrointestinal tract of the respective fish species will establish within the original host more efficiently. Here, 248 autochthonous isolates were cultured from the digestive system of three temperate flatfish species. Upon 16S rRNA gene sequencing of 195 isolates, 89.7% (n = 175) Gram-negatives belonging to the Alpha- (1.0%), Beta- (4.1%) and Gammaproteobacteria (84.6%) were identified. Candidate probiotics were further characterized using in vitro assays addressing 1) inhibition of pathogens, 2) degradation of plant derived anti-nutrient (saponin) and 3) the content of essential fatty acids (FA) and their precursors. Twelve isolates revealed an inhibition towards the common fish pathogen Tenacibaculum maritimum, seven were able to metabolize saponin as sole carbon and energy source and two isolates 012 Psychrobacter sp. and 047 Paracoccus sp. revealed remarkably high contents of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA). Furthermore, a rapid and cost-effective method to coat feed pellets revealed high viability of the supplemented probiotics over 54 d of storage at 4°C. Here, a strategy for the isolation and characterization of native probiotic candidates is presented that can easily be adapted to other farmed fish species. The simple coating procedure assures viability of probiotics and can thus be applied for the evaluation of probiotic candidates in the future.
Deoxynivalenol (DON) frequently contaminates animal feed, including fish feed used in aquaculture. This study intends to further investigate the effects of DON on carp (Cyprinus carpio L.) at concentrations representative for commercial fish feeds. Experimental feeding with 352, 619 or 953 μg DON kg−1 feed resulted in unaltered growth performance of fish during six weeks of experimentation, but increased lipid peroxidation was observed in liver, head kidney and spleen after feeding of fish with the highest DON concentration. These effects of DON were mostly reversible by two weeks of feeding the uncontaminated control diet. Histopathological scoring revealed increased liver damage in DON-treated fish, which persisted even after the recovery phase. At the highest DON concentration, significantly more fat, and consequently, increased energy content, was found in whole fish body homogenates. This suggests that DON affects nutrient metabolism in carp. Changes of lactate dehydrogenase (LDH) activity in kidneys and muscle and high lactate levels in serum indicate an effect of DON on anaerobic metabolism. Serum albumin was reduced by feeding the medium and a high dosage of DON, probably due to the ribotoxic action of DON. Thus, the present study provides evidence of the effects of DON on liver function and metabolism.
The present study investigated the effect of varying dietary protein-to-lipid ratios (P/L) on growth performance and body composition of pike perch (Sander lucioperca) fingerlings with an initial body weight of 1.35–1.40 g. Triplicate groups were reared during 56 days in recirculation systems and fed six experimental diets containing three dietary levels of lipid (90, 130, 170 g kg−1 of dry matter, d.m.) combined with two levels of dietary protein (470, 540 g kg−1 d.m.). During the experimental period the fixed feeding rates decreased from 10% to 5% of the total fish biomass per day. In contrast to previous results obtained with this species, the present study clearly demonstrated the occurrence of protein sparing effect. The best results for feed conversation ratio and specific growth rates were obtained in the treatments containing highest lipid level: diet 3 (P/L 47/17) and diet 6 (P/L 53/17). The d.m. and crude lipid contents of pike perch whole body increased in direct proportion of the dietary lipid levels. No difference was observed on crude protein and ash content among the treatments. Increasing hepatosomatic index at highest dietary lipid level suggested a limited lipid utilization of percids.
The mycotoxin zearalenone (ZEN) is frequently contaminating animal feeds including feed used in aquaculture. In the present study, the effects of dietary exposure to ZEN on carp (Cyprinus carpio L.) were investigated. ZEN at three different concentrations (low dose: 332 µg kg−1, medium dose: 621 µg kg−1 and high dose: 797 µg kg−1 final feed, respectively) was administered to juvenile carp for four weeks. Additional groups received the mycotoxin for the same time period but were fed with the uncontaminated diet for two more weeks to examine the reversibility of the ZEN effects. No effects on growth were observed during the feeding trial, but effects on haematological parameters occurred. In addition, an influence on white blood cell counts was noted whereby granulocytes and monocytes were affected in fish treated with the medium and high dose ZEN diet. In muscle samples, marginal ZEN and α-zearalenol (α-ZEL) concentrations were detected. Furthermore, the genotoxic potential of ZEN was confirmed by analysing formation of micronuclei in erythrocytes. In contrast to previous reports on other fish species, estrogenic effects measured as vitellogenin concentrations in serum samples were not increased by dietary exposure to ZEN. This is probably due to the fact that ZEN is rapidly metabolized in carp.
To produce fish of a high quality that are rich in omega-3 fatty acids (n-3 FA) and simultaneously generate more sustainable aquaculture, the combined use of phytochemicals and vegetable oils in fish feed seems to be a promising approach. Resveratrol (RV) potentially induces endogenous fatty acid synthesis, resulting in elevated n-3 FA levels in fish. RV putatively influences ∆6-desaturase, the key enzyme in FA metabolism, and serves as a ligand for PPARα, a transcription factor regulating β-oxidation. Rainbow trout (36.35 ± 0.03 g) were randomly allocated into six groups and fed diets with reduced fish oil levels (F4 = 4%, F2 = 2% and F0 = 0% of dry matter) supplemented with 0.3% (w/w) RV (F4 + RV, F2 + RV and F0 + RV). RV significantly affected FA composition in liver tissue and whole fish homogenates. 20:5n-3 (EPA) and 22:6n-3 (DHA) were significantly increased whereas precursor FA were diminished in fish fed the F2 + RV and F0 + RV diets when compared to F4 + RV and F0. RV significantly elevated ∆6-desaturase protein levels in the livers of F0 + RV fed animals. Hepatic mRNA expression of ∆6-desaturase, PPARα, and its target genes were affected by the dietary fish oil level and not by dietary RV. The results of this study indicated a potential benefit of supplementing RV in fish oil deprived diets elevating n-3 FA levels in rainbow trout.
This study evaluated a diverse range of markers of feeding stress to obtain a more precise assessment of the welfare of rainbow trout in relation to inadequate husbandry conditions. A feeding stress model based on dietary soybean meal was employed to identify suitable minimally invasive “classical” stress markers, together with molecular signatures. In a 56-day feeding experiment, rainbow trout were fed diets containing different levels of soybean meal. The impact of these different soybean meal diets on rainbow trout was assessed by water quality analyses, clinical health observations, classic growth and performance parameters, gut histopathology, blood-parameter measurements and multigene-expression profiling in RNA from whole blood. Soybean meal-induced enteritis was manifested phenotypically by an inflammatory reaction in the posterior section of the intestine and by diarrhoea in some trout. These inflammatory changes were associated with decreased supranuclear vacuolation. The haematocrit values and the levels of plasma cortisol and circulating lymphocytes in the blood were increased in trout that had consumed high amounts of SBM. Notably, the increased haematocrit depended significantly on the bodyweight of the individual trout. The transcript levels of certain genes (e.g., MAP3K1, LYG, NOD1, STAT1 and HSP90AB) emerged as potentially useful indicators in the blood of rainbow trout providing valuable information about inadequate nutrition. The expression-profiling findings provide a basis for improved, minimally invasive monitoring of feeding regimens in trout farming and may stimulate the development of practical detection devices for innovative aquaculture operations.