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.

Evaluation of graded levels of soy oil as a fish oil replacement in high soy protein feeds for juvenile red drum, Sciaenops ocellatus.

A 9-week feeding trial was conducted with juvenile red drum, Sciaenops ocellatus, to evaluate the use of soy oil as a fish oil replacement. Three primary protein sources (fishmeal - FM, soybean meal - SBM, and soy protein concentrate - SPC) were utilized with 100% fish oil (FM, SBM, SPC), 75% fish oil (SBM, SPC), or 50% fish oil (FM, SBM, SPC) as the lipid source. Traditional growth and performance metrics (specific growth rate, feed consumption, feed conversion ratio) were tracked and tissue samples (liver, muscle, plasma, adipose, and brain) were collected for gas chromatography-based fatty acid profiling. Ten lipid metabolism related genes were analyzed for potential expression differences between dietary treatments in liver and muscle tissues and whole body and fillet tissues were sampled for proximate composition analyses. Forty- four fatty acids were measured by gas chromatography-flame ionization detector (GC-FID) and evaluated with principle component analysis and ANOVA to understand the dietary influence on lipid metabolism and health. Significant differences in growth rate were observed with the SBM 50% fish oil diet outperforming the FM 100% fish oil reference diet. All other soy protein-based diets performed statistically equivalent to both FM reference diets (100% and 50% fish oil) in regard to growth, however all soy protein-based formulations had significantly lower feed conversion ratios than the fishmeal-based references (p < .001). Gene expression differences were not significant in most cases, however often trended similarly as the observed performance. Fatty acid profiles differed as a function of oil source, with no apparent influence by protein source, with C18:2n-6 (linoleic acid) being-the primary differentiator. Overall, the six soy protein, fishmeal-free formulations performed equivalently or better than the fishmeal references with up to 50% of fish oil replaced by soybean oil.

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.

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.

Metabolomics Analysis of Effects of Commercial Soy-based Protein Products in Red Drum (Sciaenops ocellatus).

We investigated the metabolic effects of four different commercial soy-based protein products on red drum fish (Sciaenops ocellatus) using nuclear magnetic resonance (NMR) spectroscopy-based metabolomics along with unsupervised principal component analysis (PCA) to evaluate metabolic profiles in liver, muscle, and plasma tissues. Specifically, during a 12 week feeding trial, juvenile red drum maintained in an indoor recirculating aquaculture system were fed four different commercially available soy formulations, containing the same amount of crude protein, and two reference diets as performance controls: a 60% soybean meal diet that had been used in a previous trial in our lab and a natural diet. Red drum liver, muscle, and plasma tissues were sampled at multiple time points to provide a more accurate snapshot of specific metabolic states during the grow-out. PCA score plots derived from NMR spectroscopy data sets showed significant differences between fish fed the natural diet and the soy-based diets, in both liver and muscle tissues. While red drum tolerated the inclusion of soy with good feed conversion ratios, a comparison to fish fed the natural diet revealed that the soy-fed fish in this study displayed a distinct metabolic signature characterized by increased protein and lipid catabolism, suggesting an energetic imbalance. Furthermore, among the soy-based formulations, one diet showed a more pronounced catabolic signature.

Acoustic monitoring indicates a correlation between calling and spawning in captive spotted seatrout (Cynoscion nebulosus).

BACKGROUND: Fish sound production is widespread throughout many families. Territorial displays and courtship are the most common reasons for fish sound production. Yet, there is still some questions on how acoustic signaling and reproduction are correlated in many sound-producing species. In the present study, our aim was to determine if a quantitative relationship exists between calling and egg deposition in captive spotted seatrout (Cynoscion nebulosus). This type of data is essential if passive acoustics is to be used to identify spawning aggregations over large spatial scales and monitor reproductive activity over annual and decadal timeframes. METHODS: Acoustic recorders (i.e., DSG-Oceans) were placed in three laboratory tanks to record underwater sound over an entire, simulated reproductive season. We enumerated the number of calls, calculated the received sound pressure level, and counted the number of eggs every morning in each tank. RESULTS: Spotted seatrout produced three distinct call types characterized as "drums," "grunts," and "staccatos." Spotted seatrout calling increased as the light cycle shifted from 13.5 to 14.5 h of light, and the temperature increased to 27.7 °C. Calling decreased once the temperature fell below 27.7 °C, and the light cycle shifted to 12 h of light. These temperature and light patterns followed the natural reproductive season observed in wild spotted seatrout in the Southeast United States. Spotted seatrout exhibited daily rhythms in calling. Acoustic signaling began once the lights turned off, and calling reached maximum activity approximately 3 h later. Eggs were released only on evenings in which spotted seatrout were calling. In all tanks, spotted seatrout were more likely to spawn when male fish called more frequently. A positive relationship between SPL and the number of eggs collected was found in Tanks 1 and 3. DISCUSSION: Our findings indicate that acoustic metrics can predict spawning potential. These findings are important because plankton tows may not accurately reflect spawning locations since egg capture is likely affected by predator activity and water currents. Instead, passive acoustics could be used to monitor spotted seatrout reproduction. Future studies can use this captive study as a model to record the estuarine soundscape precisely over long time periods to better understand how human-made stressors (e.g., climate change, noise pollution, and chemical pollutants) may affect spawning patterns.

Genetic population structure of US atlantic coastal striped bass (Morone saxatilis).

Genetic population structure of anadromous striped bass along the US Atlantic coast was analyzed using 14 neutral nuclear DNA microsatellites. Young-of-the-year and adult striped bass (n = 1114) were sampled from Hudson River, Delaware River, Chesapeake Bay, North Carolina, and South Carolina. Analyses indicated clear population structure with significant genetic differentiation between all regions. Global multilocus F ST was estimated at 0.028 (P < 0.001). Population structure followed an isolation-by-distance model and temporal sampling indicated a stable population structure more than 2 years at all locations. Significant structure was absent within Hudson River, whereas weak but significant genetic differences were observed between northern and southern samples in Chesapeake Bay. The largest and smallest effective striped bass population sizes were found in Chesapeake Bay and South Carolina, respectively. Coalescence analysis indicated that the highest historical gene flow has been between Chesapeake Bay and Hudson River populations, and that exchange has not been unidirectional. Bayesian analysis of contemporary migration indicated that Chesapeake Bay serves as a major source of migrants for Atlantic coastal regions from Albemarle Sound northward. In addition to examining population genetic structure, the data acquired during this project were capable of serving as a baseline for assigning fish with unknown origin to source region.