Investigation of the suitability of 3 insect meals as protein sources for rainbow trout (Oncorhynchus mykiss).

An in vivo trial was conducted to determine the apparent digestibility coefficients (ADCs) of insect meals for rainbow trout, Oncorhynchus mykiss. Rainbow trout (approximately 370 g ±â€…23 g, mean ±â€…SD initial weight) were stocked 25 per tank into 400-liter tanks. Fish were fed a reference diet, or 1 of 5 test diets created by blending the reference diet in a 70:30 ratio (dry-weight basis) with menhaden fish meal (MFM), 2 house cricket (Acheta domesticus) meals (cricket A and cricket B), Galleria mellonella meal, and yellow mealworm (Tenebrio molitor) meal. Diets were assigned to 3 replicate tanks of fish and fed twice daily for 14 days prior to fecal collection. Ingredients, diets, and fecal matter were analyzed in duplicate for proximate, mineral, and amino acid composition. House cricket meals were 67.3% and 69.0% protein (CP) and 16.6% and 17.1% lipid (CL), for house cricket A and B, respectively. Yellow mealworm meal contained 56.5% CP and 27.7% CL, and G. mellonella larvae meal contained 32.5% CP and 54.2% CL. Protein ADCs were 78.9 for G. mellonella larvae meal, 78.0 for yellow mealworm meal, and 76.5 for house cricket A and not different from the MFM protein ADC of 76.6, while house cricket B protein ADC was 65.8 and was significantly lower than the MFM protein ADC (F = 7.39; df = 4,14; P = 0.0049). Together, these nutritional values suggest house crickets, and yellow mealworms show promise as alternative protein sources in salmonid feeds, with the potential of G. mellonella as an alternative lipid source.

Evaluation of a hepatic biomarker of nutritional imbalance in juvenile red drum (Sciaenops ocellatus) fed 60% soybean meal-based diets using NMR-based metabolomics.

A 12-week feeding trial with juvenile red drum (Sciaenops ocellatus) fed high-soybean meal (SBM) diets was conducted to investigate a putative biomarker of nutritional imbalance, N-formimino-L-glutamate (FIGLU). Three fishmeal-free, 60% SBM pelleted diets (named B12, Fol, and Met, respectively) were tested to evaluate the effects on growth performance and tissue metabolite profiles of supplementation of vitamin B12 (0.012 mg/kg), folate (10 mg/kg), methionine (1 g/kg) respectively, above basal supplementation levels. A fourth SBM-based diet (named B12/Fol/Met) was formulated with a combination of B12, folate, and methionine to attain the above-mentioned target concentrations. A fifth 60% SBM diet (named FWS) with methionine supplementation (1 g/kg above basal supplementation levels), enriched with taurine, lysine and threonine as well as minerals, was also tested. This diet contained formulation targets and additives which have allowed for replacing fishmeal with plant proteins in rainbow trout feeds. Control diets included a fishmeal-based diet (named FM), an unsupplemented basal 60% SBM diet (named SBM60), and a "natural" diet (named N) made up of equal parts of fish (cigar minnows), squid and shrimp as a positive reference for growth performance. Formulated feeds contained approximately 37% total crude protein, approximately 14% total crude lipid and were energetically balanced. Standard growth performance metrics were measured, and tissues (liver, muscle) were collected at week 12 to evaluate diet-induced metabolic changes using nuclear magnetic resonance (NMR)-based metabolomics. Our results show that the FWS diet outperformed all other SBM diets and the FM diet under all performance metrics (P < 0.05). FIGLU was not detected in fish fed the N diet but was detected in those fed the SBM diets and the FM diet. Fish fed the FWS diet and the Met diet showed lower hepatic levels of FIGLU compared with the other SBM-based diets (P < 0.05), suggesting that among the different supplementation regimes, methionine supplementation was associated with lower FIGLU levels. The FWS diet produced tissue metabolite profiles that were more similar to those of fish fed the N diet. Based on our results, the FWS diet constitutes a promising SBM-based alternative diet to fishmeal for red drum.

Validation of a microwave energy meter to non-lethally estimate energetic reserves in adult sturgeon.

Whole-body (WB) energetic reserves influence fish survival, growth, and reproduction but are typically quantified using lethal methods (i.e. proximate analyses) or interpreted through body condition indices. Energetic reserves can impact population dynamics through influences on growth rates, age-at-first-reproductive-maturity, and spawning periodicity at the individual-fish level, especially in long-lived sturgeon species. Therefore, a non-lethal tool to track the energetic reserves of endangered sturgeon populations could inform adaptive management and further our understanding of the sturgeon's biology. The Distell Fatmeter is a microwave energy meter that has been validated to non-lethally estimate energetic reserves in some fish species, but never successfully for sturgeon. Here, stepwise linear regressions were applied to test commonly monitored body metrics and Fatmeter measurements at nine different anatomical sites on captive adult pallid sturgeon (Scaphirhynchus albus; total length of 790-1015 mm; WB lipid of 13.9-33.3%) compared with WB lipid and energy content determined by proximate analyses. Fatmeter measurements alone explained approximately 70% of the variation in WB energetic reserves, which outperformed models considering body metrics alone by a margin of approximately 20%. The top-ranked models based on AICc score (second-order Akaike Information Criterion) included a combination of body metrics and Fatmeter measurements and accounted for up to 76% of the variation in WB lipid and energy. We recommend the incorporation of Fatmeter measurements at a single site located dorsally to the lateral scutes at the posterior end of the fish above the pelvic fins (U-P) into conservation monitoring programs for adult pallid sturgeon (total length [TL] ≥ 790 mm; fork length [FL] ≥ 715 mm) and the cautious application of Fatmeter measurements for sturgeon between 435 and 790 mm TL (375-715 mm FL). Measurements at this U-P site combined with body mass explained approximately 75% of the variation in WB lipid and energy.

Investigation of graded levels of soybean meal diets for red drum, Sciaenops ocellatus, using quantitative PCR derived biomarkers.

A twelve-week feeding trial was conducted to examine potential metabolic and gene expression changes that occur in juvenile red drum, Sciaenops ocellatus, fed diets with increasing soybean meal inclusion. Significant reduction in fish performance characteristics (feed consumption, weight gain, final weight) was observed within the soybean meal based diets as soybean meal level increased (R, linear regression); however, all soybean meal based diets performed statistically equivalent in regards to performance characteristics (weight gain, feed conversion ratio, condition factor, etc.) to a commercial (45% crude protein and 16% crude lipid) reference diet (R, ANOVA). To better understand the underlying physiological responses and metabolic changes driving performance differences, traditional aquaculture metrics were paired with high throughput -omics techniques. Nuclear magnetic resonance (NMR) spectroscopy-based metabolomics data and RNA transcript abundance differences observed in liver tissue were utilized to select multiple sets of genes to target with quantitative polymerase chain reaction (qPCR), both for pathway activity validation and as rapid and accessible biomarkers of performance as a result of soybean meal. Genes selected based on the metabolic pathways most affected by soybean meal level corroborate the metabolite profile and performance data indicating an increase in gluconeogenic precursor production as soybean meal increased. The metabolomic and gene expression tools utilized in our study present a novel framework for diet and fish health evaluation that may provide more rapid and improved techniques for evaluating dietary manipulations and improving production of juvenile fish on alternative feeds.

Effects of high-soy diet on S100 gene expression in liver and intestine of rainbow trout (Oncorhynchus mykiss).

The current study examines expression of S100 genes, a group of calcium-sensing proteins poorly characterized in fishes. In mammals, these proteins are known to play roles beyond calcium-signaling, including mediation of inflammatory processes. Some S100 proteins also serve as biomarkers for a variety of autoinflammatory conditions. It is well known that salmonids exhibit varying degrees of intestinal enteritis when exposed to alternative feed ingredients containing antinutritional factors, with soybean meal (SBM) being one of the best characterized. The etiology of soy-caused distal enteritis isn't entirely understood but displays similar histopathological alterations to the gut observed in human mucosal inflammatory bowel diseases. We sought to determine if teleost S100 genes show a concomitant response like that observed in mammals, utilizing rainbow trout fed high-soy diets as a model for intestinal inflammation. We examined expression of fourteen known salmonid S100 genes in the liver, first segment of the mid-intestine (proximal intestine), and second segment of the mid-intestine (distal intestine). After 12 weeks on a high-soy diet containing 40% SBM, we observed upregulation of several S100 genes in the distal intestine (S100I2, A10a, V1, V2, and W), no changes in the proximal intestine, and downregulation of S100V2 in the liver. Overall, our results provide further knowledge of the expression of S100 genes and provide targets for future research regarding inflammatory processes in the rainbow trout gut.

Investigation of graded-level soybean meal diets in red drum (Sciaenops ocellatus) using NMR-based metabolomics analysis.

We investigated changes in the metabolome in juvenile red drum (Sciaenops ocellatus) induced by increasing amounts of soybean meal (0% to 60%) in extruded, fishmeal-free diets using a nuclear magnetic resonance spectroscopy (NMR)-based metabolomics approach in a 12-week feeding trial. All of the diets were composed of ≈40% total crude protein, ≈11% total crude lipid and were energetically balanced. A fishmeal-containing, commercial extruded diet was used as a control diet throughout the trial. Each week, liver, muscle, intestine and plasma samples were collected and analyzed by NMR to provide a "snapshot" of the metabolome at different time points. Results indicate significant time-dependence of the metabolic profiles in various tissues with stable metabolomic profiles forming after about 9-weeks on the experimental diets. We identify a previously unexploited biomarker of potential dietary stress (N­formimino­l­glutamate (FIGLU)) in the fish that may prove to be useful for optimization of alternative diet formulations.

Water system is a controlling variable modulating bacterial diversity of gastrointestinal tract and performance in rainbow trout.

A two-phase feeding study evaluating performance of rainbow trout and comparing luminal and mucosal gastrointestinal tract (GIT) bacterial community compositions when fed two alternative protein diets in two rearing systems was conducted. Alternative protein diets (animal protein and plant protein diets) balanced with crystalline amino acids: lysine, methionine and threonine or unbalanced, were fed to rainbow trout in two separate water systems (recirculating (RR) and flow-through (FF)) for a period of 16 weeks. The four diets, each contained 38% digestible protein and 20% fats, were fed to rainbow trout with an average weight of 12.02 ± 0.61 g, and sorted at 30 fish/tank and 12 tanks per dietary treatment. Phase 1 lasted for 8 weeks after which fish from each tank were randomly divided, with one-half moved to new tanks of the opposing system (i.e. from RR to FF and vice versa). The remaining halves were retained in their initial tank and system, and fed their original diets for another 8 weeks (phase 2). After the 16th week, 3 fish/tank were sampled for each of proximate analysis, body indexes and 16S rRNA analysis of GIT microbiota. Fish weight (P = 0.0008, P = 0.0030, P<0.0010) and body fat (P = 0.0008, P = 0.0041, P = 0.0177) were significantly affected by diet, diet quality (balanced or unbalanced) and system, respectively. Feed intake (P = 0.0008) and body energy (P<0.0010) were altered by system. Body indexes were not affected by dietary treatment and water systems. Compositional dissimilarities existed between samples from the rearing water and GIT locations (ANOSIM: (R = 0.29, P = 0.0010), PERMANOVA: R = 0.39, P = 0.0010), but not in dietary samples (ANOSIM: R = 0.004, P = 0.3140, PERMANOVA: R = 0.008, P = 0.4540). Bacteria were predominantly from the phyla Proteobacteria, Firmicutes and Bacteroidetes. Their abundance differed with more dissimilarity in the luminal samples (ANOSIM: R = 0.40, P = 0.0010, PERMANOVA: R = 0.56, P = 0.0010) than those from the mucosal intestine (ANOSIM: R = 0.37, P = 0.0010, PERMANOVA: R = 0.41, P = 0.0010). Bacteria generally associated with carbohydrate and certain amino acids metabolism were observed in the mucosal intestine while rearing water appeared to serve as the main route of colonization of Aeromonas and Acinetobacter in the rainbow trout.

Effect of fasting on body composition and responses to stress in sunshine bass.

The integrated responses of the hormonal regulation of growth and stress in sunshine bass (Morone chrysops X Morone saxatilis) as regulated by feed deprivation were investigated. Groups of fish were fed 1.5% of the body weight per day or offered no feed for 4weeks. Another group of fish was not fed for 3weeks and feed was offered during the fourth week. Fish in each group were sampled immediately before or after a 15-min low water confinement stressor after each week of the experiment. Liver mass and liver glycogen content were decreased after one week of fasting and remained low until the end of the study. However, both recovered after a week of refeeding. Intraperitoneal fat was significantly lower after two weeks of fasting and did not recover after a week of refeeding. None of these components were affected by confinement stress. Plasma glucose in unstressed fish was generally unaffected by fasting or refeeding; however, plasma glucose increased after confinement stress in fed but not in fasted fish. The cortisol stress response was unaltered by fasting and remained robust. Plasma IGF-I generally decreased in fasted fish but was not significantly lower than fed fish until the fourth week. A week of refeeding did not restore plasma IGF-I concentrations. Plasma IGF-I concentrations were higher in confinement stressed fed fish after two and four weeks but were unchanged in the fourth week. There was no change in the plasma IGF-I concentrations in fasted or refed fish due to the stress. Liver weight and liver glycogen were essentially depleted after 2weeks of fasting. The reduction of liver glycogen greatly reduced the glucose response to stress; however, the cortisol stress response was maintained for at least four weeks of fasting. Intraperitoneal fat was decreased very little after 4weeks of fasting. Plasma IGF-I concentrations were reduced only after 3weeks of fasting.

Determination of relative protein degradation activity at different life stages in rainbow trout (Oncorhynchus mykiss).

Rainbow trout were reared from 5 g to approximately 400 g on a diet formulated to supply the required protein from either fishmeal or plant proteins. The fish were sampled at every weight doubling and liver and muscle samples were obtained. From these tissue samples RNA and protein were isolated and analyzed for the expression of a number of muscle regulatory and protein degradation genes and enzymatic activity for proteins involved in the caspase, calpain, and ubiquitin-proteasome pathways for protein proteolysis. Only MyoD2 showed significant differences in expression between the two diets, while no significant changes over the course of the experiment were determined for MyoD2 or the other muscle factors. For the degradation genes significant changes in expression were determined for calpain1 and calpastatin. Calpastatin also showed a significant increase in expression over the course of the experiment in the muscle of fish fed a fishmeal diet and significant decrease in expression in the liver of fish fed the fishmeal based diet. Differences in proteasome enzyme activity were found between diets in the liver and muscle of fish and for caspase-3 activity in muscle. Significant changes in activity over the course of the experiment were noted for proteasome and calpain activity in the liver and muscle. These findings suggest that diets replacing fishmeal with plant material can have some effects on protein turnover in muscle and that some degradation pathways are differentially regulated during the growth of rainbow trout.