Assessment of dietary yeast-based additives for cultured catfish and tilapia health.

Channel catfish (Ictalurus punctatus) and Nile tilapia (Oreochromis niloticus) are two aquaculture species of great importance. Intensive production is often hindered by poor growth performance and disease mortality. The aim of this study was to evaluate the potential of a commercial fermented yeast product, DVAQUA, on channel catfish and Nile tilapia growth performance metrics and disease resistance. Channel catfish and Nile tilapia were fed practical diets supplemented with 0%, 0.1% or 0.4% of DVAQUA over approximately 2-month feeding periods in recirculation aquaculture systems. To assess the potential of the postbiotic against common aquaculture pathogens, juvenile catfish were subsequently challenged by immersion with Edwardsiella ictaluri S97-773 or virulent Aeromonas hydrophila ML09-119. Nile tilapia juveniles were challenged by injection with Streptococcus iniae ARS-98-60. Serum lysozyme activity, blood chemistry and growth metrics were measured at the end of the feeding period, but no differences were observed across the different metrics, except for survival. For the pathogen challenges, there were no differences in endpoint mortality for channel catfish with either pathogen (p > .05). In contrast, Nile tilapia survivability to S. iniae infection increased proportionally to the inclusion of DVAQUA (p = .005). Changes to sera lysozyme activity were also noted in the tilapia trial, with a reduction of activity in the fish fed the 0.4% DVAQUA diet compared to the control diet (p = .031). Expression profiles of proinflammatory genes and antibodies were also found to be modulated in channel catfish fed the postbiotic, indicating some degree of protective response. These results suggest that this postbiotic may be beneficial in protecting Nile tilapia against S. iniae infection by influencing immune parameters and additional research is needed to evaluate the potential of this DVAQUA for improving catfish health and disease control.

Investigation of dietary exogenous protease and humic substance on growth, disease resistance to Flavobacterium covae and immune responses in juvenile channel catfish (Ictalurus punctatus).

Many bacterial pathogens impact the US catfish industry, and disease control can be challenging for producers. Columnaris disease in channel catfish, Ictalurus punctatus, is primarily caused by Flavobacterium covae (formerly F. columnare). Immunostimulants may enhance nonspecific immune responses and offer an alternative to antibiotic treatments in catfish. Furthermore, dietary protein sources and inclusions are also essential to fish health and nutrition and may enhance overall fish performance in pond culture. The current project evaluated two immunostimulants: a protease complex (PC) and a humic substance (HS) derived from a reed-sedge peat product. A 60-day trial examined the effects of five diets on growth performance, immune response and resistance to experimental F. covae infection in channel catfish. Diets included a high-quality fishmeal diet (32%; CF32), a high-protein soy-based diet (32%; C32), a low-protein soy-based diet (28%; C28; predominately used in industry), a low-protein soy diet supplemented with C28 + PC at 175 g metric ton-1 and C28 + HS in a low-protein diet at 23 g metric ton-1 . Following feeding for 60 d, juvenile channel catfish were sampled for growth performance and baseline health indicators (n = 3; body mucus, blood for sera, kidney and spleen). A subset of fish was then subjected to an immersion-based in vivo challenge trial with F. covae (ALG-00-530; 106 CFU mL-1 exposure). At 60d post-initiation, there were no dietary differences in the relative growth rate (p = .063) or thermal growth coefficient (p = .055), but the 32% diets generally appeared to perform best. Post-challenge, the C32 group's mortality was higher than the C28 + PC (p = .006) and C28 + HS diets (p = .005). Although not significant, the C28 and CF32 groups also demonstrated higher mortality compared to both PC and HS diets. Sera lysozyme concentration was found to increase following pathogen challenge (p < .001) and in comparison with mock-challenged catfish (p < .001). Elevated expression levels of proinflammatory cytokines (il-1ß, il-8, tnf-α and tgf-ß) were observed at trial midpoint and post-infection when compared to 60d. The C28 treatment was found to have lower tnf-α expression than the C28 + PC (p = .042) and C28 + HS (p = .042) groups following exposure to F. covae. These challenge data suggest that the immunostimulants (PC and HS) in plant-based protein may be beneficial in protecting against F. covae when offered in low-protein channel catfish diets.

Probiotics enhance resistance to Streptococcus iniae in Nile tilapia (Oreochromis niloticus) reared in biofloc systems.

Biofloc technology is a rearing technique that maintains desired water quality by manipulating carbon and nitrogen and their inherent mixture of organic matter and microbes. Beneficial microorganisms in biofloc systems produce bioactive metabolites that may deter the growth of pathogenic microbes. As little is known about the interaction of biofloc systems and the addition of probiotics, this study focused on this integration to manipulate the microbial community and its interactions within biofloc systems. The present study evaluated two probiotics (B. velezensis AP193 and BiOWiSH FeedBuilder Syn 3) for use in Nile tilapia (Oreochromis niloticus) culture in a biofloc system. Nine independent 3785 L circular tanks were stocked with 120 juveniles (71.4 ± 4.4 g). Tilapia were fed for 16 weeks and randomly assigned three diets: a commercial control diet or a commercial diet top-coated with either AP193 or BiOWiSH FeedBuilder Syn3. At 14 weeks, the fish were challenged with a low dose of Streptococcus iniae (ARS-98-60, 7.2 × 107 CFU mL-1 , via intraperitoneal injection) in a common garden experimental design. At 16 weeks, the fish were challenged with a high dose of S. iniae (6.6 × 108 CFU mL-1 ) in the same manner. At the end of each challenge trial, cumulative per cent mortality, lysozyme activity and expression of 4 genes (il-1ß, il6, il8 and tnfα) from the spleen were measured. In both challenges, the mortalities of the probiotic-fed groups were significantly lower (p < .05) than in the control diet. Although there were some strong trends, probiotic applications did not result in significant immune gene expression changes related to diet during the pre-trial period and following exposure to S. iniae. Nonetheless, overall il6 expression was lower in fish challenged with a high dose of ARS-98-60, while tnfα expression was lower in fish subjected to a lower pathogen dose. Study findings demonstrate the applicability of probiotics as a dietary supplement for tilapia reared in biofloc systems.

Transcriptomic profiles of Florida pompano (Trachinotus carolinus) gill following infection by the ectoparasite Amyloodiniumocellatum.

The dinoflagellate Amyloodinium ocellatum is an important pathogenic parasite infecting cultured marine and brackish water fishes worldwide. This includes cultured Florida pompano (Trachinotus carolinus), which is one of the most desirable marine food fish with high economic value in the USA. A. ocellatum infects fish gills and causes tissue damage, increased respiratory rate, reduced appetite, and mortality, especially in closed aquaculture systems. This study mimicked the natural infection of A. ocellatum in cultured pompano and conducted a transcriptomic comparison of gene expression in the gills of control and A. ocellatum infected fish to explore the molecular mechanisms of infection. RNA-seq data revealed 604 differentially expressed genes in the infected fish gills. The immunoglobulin genes (including IgM/T) augmentation and IL1 inflammation suppression were detected after infection. Genes involved in reactive oxygen species mediating parasite killing were also highly induced. However, excessive oxidants have been linked to oxidative tissue damage and apoptosis. Correspondingly, widespread down-regulation of collagen genes and growth factor deprivation indicated impaired tissue repair, and meanwhile the key executor of apoptosis, caspase-3 was highly expressed (25.02-fold) in infected fish. The infection also influenced the respiratory gas sensing and transport genes and established hypoxic conditions in the gill tissue. Additionally, food intake and lipid metabolism were also affected. Our work provides the transcriptome sequencing of Florida pompano and provides key insights into the acute pathogenesis of A. ocellatum. This information can be utilized for designing optimal disease surveillance strategies, future selection for host resistance, and development of novel therapeutic measures.

Effect of salinity on growth, survival, and serum osmolality of red snapper, Lutjanus campechanus.

Three trials were conducted to evaluate the performances of red snapper, Lutjanus campechanus, in low salinities. The median lethal concentration (96 h LC50) of salinity was determined by trimmed Spearman-Karber method using survival data of fish (18.9 ± 0.2 g) collected after 96 h from acclimation to 2, 4, 8, and 32 ppt salinities in 800 L tanks (n = 3), while the serum osmolality of fish (74.1 ± 3.9 g) was determined after 48 h from acclimation to 6, 8, 16, 24, and 32 ppt salinities in 150 L tanks (n = 3). The growth trial was conducted for 6 weeks in 800 L tanks to determine the growth and survival of fish (18.8 ± 0.2 g) at 8 ppt salinity compared to the control (32 ppt salinity). At the conclusion, the isosmotic point of fish was estimated as 357.2 mmol/kg (correspond to 11.0 ppt salinity), while the 96 h LC50 was estimated as 5.65 ppt salinity. No significant differences were noted for survival and FCR of fish reared in 8 and 32 ppt salinities. However, growth was significantly lower in fish reared in 8 ppt salinity compared to the fish reared in 32 ppt salinity. The reduced growth could be, at least partially, due to the increased osmoregulatory energy expenditure at lower salinities.

Replacement of fish meal with Bacillus pumillus SE5 and Pseudozyma aphidis ZR1 fermented soybean meal in diets for Japanese seabass (Lateolabrax japonicus).

This study examined the effects of replacing fish meal (FM) with three different types of soybean meal (SM) including untreated SM, Bacillus pumillus SE5 (BP) fermented SM (BPFSM) and Pseudozyma aphidis ZR1 (PA) fermented SM (PAFSM) in diets for Japanese seabass (Lateolabrax japonicus). A basal diet was formulated using FM (FM diet), and six other diets were produced by substituting 40 or 80% of FM with SM, BPFSM or PAFSM (SM40, SM80, BPFSM40, BPFSM80, PAFSM40 and PAFSM80 diets). Each diet was fed to triplicate groups of fish (7.14 ±â€¯0.05 g) twice daily for eight weeks. Replacing 40% of FM with SM sources did not significantly influence growth (P > 0.05), while increasing the substitution level to 80% led to reduced growth rates (P < 0.05). The groups received SM80 and PAFSM80 diets showed significantly higher feed conversion ratio and lower protein digestibility than FM group. Furthermore, notably lower dry matter digestibility was detected in SM80 group. Remarkably lower serum total antioxidant capacity was found in the SM80 group, and catalase activity did not significantly differ between FM and BPFSM40 groups. Serum malondialdehyde concentration was enhanced by increasing FM replacement level and the highest value was observed in the SM80 fed fish. FM and PAFSM40 groups showed significantly higher lysozyme activity than the SM80 group. Fish fed the BPFSM40 diet exhibited the highest complement C3 activity and the lowest value was observed in the SM80 group. Expression of lysozyme gene in spleen was down-regulated in the SM80 group, and no significant difference in expression of C3 gene was found among FM, BPFSM40 and PAFSM40 groups. Digestive enzymes activity and gut morphology were significantly influenced by FM replacement. Expression of HSP70 and pro-inflammatory genes including TNF-α and IL-1ß were up-regulated by FM replacement and relatively lower expression levels were found by using fermented SM. An opposite trend was observed for the anti-inflammatory TGF-ß gene expression. Serum d-lactate concentration was significantly increased by replacing 80% of FM with any of the SM sources. These findings indicated that using fermented SM, particularly BPFSM, beneficially influences feed utilization, antioxidant capacity, innate immunity and gut health in juvenile Japanese seabass.

Impact of feed additives on surface mucosal health and columnaris susceptibility in channel catfish fingerlings, Ictalurus punctatus.

One of the highest priority areas for improvement in aquaculture is the development of dietary additives and formulations which provide for complete mucosal health and protection of fish raised in intensive systems. Far greater attention has been paid to dietary impact on gut health than to protective effects at other mucosal surfaces such as skin and gill. These exterior surfaces, however, are important primary targets for pathogen attachment and invasion. Flavobacterium columnare, the causative agent of columnaris disease, is among the most prevalent of all freshwater disease-causing bacteria, impacting global aquaculture of catfish, salmonids, baitfish and aquaria-trade species among others. This study evaluated whether the feeding of a standard catfish diet supplemented with Alltech dietary additives Actigen(®), a concentrated source of yeast cell wall-derived material and/or Allzyme(®) SSF, a fermented strain of Aspergillus niger, could offer protection against F. columnare mortality. A nine-week feeding trial of channel catfish fingerlings with basal diet (B), B + Allzyme(®) SSF, B + Actigen(®) and B + Actigen(®)+Allzyme(®) SSF revealed good growth in all conditions (FCR < 1.0), but no statistical differences in growth between the treatments were found. At nine weeks, based on pre-challenge trial results, basal, B + Actigen(®), and B + Allzyme(®) SSF groups of fish were selected for further challenges with F. columnare. Replicated challenge with a virulent F. columnare strain, revealed significantly longer median days to death in B + Allzyme(®) SSF and B + Actigen(®) when compared with the basal diet (P < 0.05) and significantly higher survival following the eight day challenge period in B + Actigen(®) when compared with the other two diets (P < 0.05). Given the superior protection provided by the B + Actigen(®) diet, we carried out transcriptomic comparison of gene expression of fish fed that diet and the basal diet before and after columnaris challenge using high-throughput RNA-seq. Pathway and enrichment analyses revealed changes in mannose receptor DEC205 and IL4 signaling at 0 h (prior to challenge) which likely explain a dramatic divergence in expression profiles between the two diets soon after pathogen challenge (8 h). Dietary mannose priming resulted in reduced expression of inflammatory cytokines, shifting response patterns instead to favor resolution and repair. Our results indicate that prebiotic dietary additives may provide protection extending beyond the gut to surface mucosa.

Practical diets for California yellowtail, Seriola dorsalis: Use of advanced soybean meal products on growth performance, body composition, intestinal morphology, and immune gene expression.

California yellowtail (CYT), Seriola dorsalis, is a promising candidate for aquaculture due to its rapid growth and high-quality flesh, particularly in markets like Japan, Australia, China, and the United States. Soy protein has shown success as a replacement for marine protein sources in CYT diets, reducing fishmeal levels, though concerns about potential intestinal inflammation persist with the inclusion of solvent-extracted soybean meal. To address this, processing strategies like fractionation, enzymatic treatment, heat treatment, and microbial fermentation have been employed to mitigate the negative impacts of soybean meal on fish nutrition and immune systems. This study focuses on optimizing soybean meal inclusion levels by incorporating advanced soy variants into CYT diets. The eight-week feeding trial, conducted in a recirculation system, featured six diets with sequential inclusion levels (0, 50, 100%) of high protein low oligosaccharide soybean meal (Bright Day, Benson Hill, St Louis, MO) and enzyme-treated soybean meal (HP 300, Hamlet Protein Inc., Findlay, OH), replacing solvent-extracted soybean. The study compares these formulations against a soy-free animal protein-based diet. At the end of the trial, fish were sampled for growth performance, body proximate composition, intestinal morphology, and immune response from gut samples. Results showed consistent FCR (P = 0.775), weight gain (P = 0.242), and high survival rate (99.4 ± 0.5%) among dietary treatments (P>0.05). Histological evaluations revealed no gut inflammation and gene expression analysis demonstrated no significant variations in immune, physiological, and digestive markers apn (P = 0.687), mga (P = 0.397), gpx1 (P = 0.279), atpase (P = 0.590), il1ß (P = 0.659). The study concludes that incorporating advanced soybean meal products, replacing up to 20% of fishmeal does not negatively affect CYT's growth and intestinal health. This suggests that all three soy sources, contributing 35% of total protein (15.4 g 100 g-1 diet), can be included in practical diets without compromising CYT's intestinal integrity or growth. These findings have positive implications for the commercial production of CYT and future research on the incorporation of plant-based proteins in aquaculture diets.

Turnovers of Sex-Determining Mutation in the Golden Pompano and Related Species Provide Insights into Microevolution of Undifferentiated Sex Chromosome.

The suppression of recombination is considered a hallmark of sex chromosome evolution. However, previous research has identified undifferentiated sex chromosomes and sex determination by single SNP in the greater amberjack (Seriola dumerili). We observed the same phenomena in the golden pompano (Trachinotus ovatus) of the same family Carangidae and discovered a different sex-determining SNP within the same gene Hsd17b1. We propose an evolutionary model elucidating the turnover of sex-determining mutations by highlighting the contrasting dynamics between purifying selection, responsible for maintaining W-linked Hsd17b1, and neutral evolution, which drives Z-linked Hsd17b1. Additionally, sporadic loss-of-function mutations in W-linked Hsd17b1 contribute to the conversion of W chromosomes into Z chromosomes. This model was directly supported by simulations, closely related species, and indirectly by zebrafish mutants. These findings shed new light on the early stages of sex chromosome evolution.

Sources of variation in elemental profiles of whiteleg shrimp (Litopenaeus vannamei) and their potential effects on the accuracy of discriminant analysis.

BACKGROUND: Elemental profiling is a tool that has been proposed to improve the traceability of seafood products. Small sources of variation can affect the outcome of elemental profiling and therefore pose to lower the overall accuracy of analyses. Here, we investigate two potential sources of variation through three experiments: laboratory variation (intra-, interlaboratory variation, and tissue matrix) and tissue variation. METHODS AND RESULTS: Samples of whitleleg shrimp (Litopenaeus vannamei) were obtained from 20 farms in Ecuador and two farms in Alabama to be analyzed. In the first experiment of the study, samples from Ecuador were analyzed at three different laboratories and compared. Two out of the five elements reported were statistically different across the three laboratories (Cu and Se). In the second experiment, the effect of tissue matrix (ground vs whole tissue during acid digestion) was investigated. Altogether, five out of 29 elements analyzed were statistically different. In the third experiment, samples from two farms in Alabama were analyzed to understand the variation in element concentrations in different tissues (head on shell on (HOSO), headless shell on (HLSO), headless peeled (PLD) and headless peeled and deveined (PLDV)). Elemental concentrations varied across tissues, and patterns in elemental concentrations were site specific. The samples from the two farms were analyzed with a Random Forest classification model to site x tissue groupings with 94% accuracy. CONCLUSION: The result of this study highlights the following: 1. Consistency in laboratory analysis important in studies that involve element concentrations, as minor differences in methodology can propagate as significant differences in results. 2. In shrimp, elements are compartmentalized in different tissues and elemental profiling should consistently use the same type of tissue.

A preliminary survey of antibiotic residues in frozen shrimp from retail stores in the United States.

Shrimp are an important and valuable commodity for aquaculture that are widely traded internationally. Widespread antibiotic use has been documented in shrimp farming and is a common source of criticism of aquaculture products. Additionally, previous reports have found some evidence of antibiotic residues in shrimp samples obtained from retail stores in the United States, which is a concern for consumers. To further understand the prevalence of antibiotics in retail shrimp in the United States, shrimp samples obtained from grocery stores across 16 states were analyzed for 74 antibiotic compounds/metabolites at a commercial laboratory. 68 samples were analyzed for a multiclass antibiotic panel which included 66 antibiotics while a subset of 15 samples were analyzed for ß-lactam antibiotics, Nitrofurans, and Oxytetracycline. Samples were obtained that were labeled as being from major production countries, including India, Indonesia, Thailand, and Vietnam. No detectable antibiotic residues were found in this survey in any samples. This is contrary to previous findings in frozen shrimp analyzed for antibiotics, which typically report low levels of the prevalence of antibiotics.

Growth, serum biochemical parameters, salinity tolerance and antioxidant enzyme activity of rainbow trout (Oncorhynchus mykiss) in response to dietary taurine levels.

This study evaluated the effect of dietary taurine levels on growth, serum biochemical parameters, salinity adaptability, and antioxidant activity of rainbow trout (Oncorhynchus mykiss). Four diets were formulated with taurine supplements at 0, 0.5, 1, and 2% w/v (abbreviated as T0, T0.5, T1, and T2, respectively). Rainbow trouts (initial weight of 80.09 ± 4.72 g) were stocked in tanks (180 L capacity), and were fed these diets for six weeks and subsequently underwent salinity acclimation. Physiological indicators were determined before salinity acclimation at 1, 4, 7, and 14 days afterwards. Results showed that there were no significant differences in growth performance (final mean weight ranged from 182.35 g to 198.48 g; percent weight gain was between 127.68% and 147.92%) of rainbow trout in freshwater stage, but dietary taurine supplement significantly increased serum-free taurine content. After entering seawater, the Na+-K+-ATPase activity of T2 group returned to its freshwater levels, and the serum cortisol content was significantly higher than T0 and T0.5 groups. At the end of this experiment, the liver superoxide dismutase activity in the T0 and T0.5 groups was significantly lower than in the T1 and T2 groups, and the liver catalase in the T0 group was the lowest whereas that in the T2 group was the highest. Muscle malondialdehyde content was the highest in the T0 group, and the lowest in the T2 group. Based on the results of this study, supplement of dietary taurine (0.5-2%) enhanced the salinity tolerance in rainbow trout, which increased with the higher taurine concentration.

Trace element concentrations in white leg shrimp Litopenaeus vannamei from retail stores in the EU, UK, and USA and the ability to discern country of origin with classification models.

Shrimp are a globally traded aquaculture commodity that accounts for a large proportion of the monetary value of aquaculture. There are concerns among consumers about seafood labeling fraud and environmental sustainability. Therefore, the geographic origin of shrimp from retail stores was investigated with trace element profiling. 94 shrimp samples were collected from grocery stores across the USA, UK, and EU in 70 different grocery stores. The results of 24 elements are reported. Shrimp samples were from Thailand, India, Vietnam, Indonesia, and Ecuador were shown to have 15 elements that were statistically different across labeled country of origin, with Ecuador having unique post hoc group membership in 5 of the elements. Based on a classification procedure, shrimp were classified to labeled country of origin with an overall accuracy of 71.2%. Overall, the results suggest that elemental profiling could be a traceability tool for classifying samples of shrimp from retail stores.

Branchial carbonic anhydrase activity and ninhydrin positive substances in the Pacific white shrimp, Litopenaeus vannamei, acclimated to low and high salinities.

The Pacific white shrimp, Litopenaeus vannamei, acclimated to 30 ppt salinity, was transferred to either low (15 and 5 ppt), or high (45 ppt) salinity for 7 days. Hemolymph osmolality, branchial carbonic anhydrase activity, and total ninhydrin-positive substances (TNPS) in abdominal muscle were then measured for each condition. Hemolymph osmotic concentration was regulated slightly below ambient water osmolality in shrimp acclimated to 30 ppt. At 15 and 5 ppt, shrimp were strong hyper-osmotic regulators, maintaining hemolymph osmolality between 200 and 400 mOsm above ambient. Shrimp acclimated to 30 ppt and transferred to 45 ppt salinity were strong hypo-osmotic and hypo-ionic regulators, maintaining hemolymph osmolality over 400 mOsm below ambient. Branchial carbonic anhydrase (CA) activity was low (approximately 100 micromol CO(2) mg protein(-1) min(-1)) and uniform across all 8 gills in shrimp acclimated to 30 ppt, but CA activity increased in all gills after exposure to both low and high salinities. Anterior gills had the largest increases in CA activity, and levels of increase were approximately the same for low and high salinity exposure. Branchial CA induction appears to be functionally important in both hyper- and hypo-osmotic regulations of hemolymph osmotic concentrations. Abdominal muscle TNPS made up between 19 and 38% of the total intracellular osmotic concentration in shrimp acclimated to 5, 15, and 30 ppt. TNPS levels did not change across this salinity range, over which hemolymph osmotic concentrations were tightly regulated. At 45 ppt, hemolymph osmolality increased, and muscle TNPS also increased, presumably to counteract intracellular water loss and restore cell volume. L. vannamei appears to employ mechanisms of both extracellular osmoregulation and intracellular volume regulation as the basis of its euryhalinity.