Assessment of Safety, Effects, and Muscle-Specific Accumulation of Dietary Butylated Hydroxytoluene (BHT) in Paralichthys olivaceus.

Butylated hydroxytoluene (BHT) is a commonly used antioxidant added to animal/fish feed to limit lipid autoxidation and peroxidation. Although there have been reviews and reports of BHT toxicity in animals, limited information is available with respect to the toxic effects and accumulation of BHT due to oral exposure in aquaculture species. Therefore, 120 days of feeding trial was conducted to evaluate the effects of dietary BHT on the marine fish olive flounder Paralichthys olivaceus. Graded levels of BHT were added to the basal diet in increments of 0, 10, 20, 40, 80, and 160 mg BHT/kg, corresponding to 0 (BHT0), 11 (BHT11), 19 (BHT19), 35 (BHT35), 85 (BHT85), and 121 (BHT121) mg BHT/kg diets, respectively. Fish with an average weight of 77.5 ± 0.3 g (mean ± SD) were fed one of the six experimental diets in triplicate groups. Growth performance, feed utilization, and survival rate were not significantly affected by the dietary BHT levels among all experimental groups, whereas BHT concentration in the muscle tissue was found to increase in a dose-dependent manner up to 60 days of the experimental period. Thereafter, BHT accumulation in muscle tissue showed a declining trend among all treatment groups. Furthermore, the whole-body proximate composition, nonspecific immune responses, and hematological parameters (except triglycerides) were not significantly influenced by the dietary levels of BHT. Blood triglyceride content was significantly higher in fish fed the BHT-free diet compared to all other treatment groups. Thus, this study demonstrates that dietary BHT (up to 121 mg/kg) is a safe and effective antioxidant without exhibiting any adverse effects on the growth performance, body composition, and immune responses in the marine fish olive flounder, P. olivaceus.

Partial Substitution of Fish Oil with Microalgae (Schizochytrium sp.) Can Improve Growth Performance, Nonspecific Immunity and Disease Resistance in Rainbow Trout, Oncorhynchus mykiss.

The price of fish oil has reached a historical peak due to a consistent downward production trend, and therefore, the search for sustainable alternative sources has received great attention. This research was conducted to evaluate dietary micro-algae, Schizochytrium sp. (SC) as fish oil (FO) replacer in rainbow trout, Oncorhynchus mykiss. In the first trial, apparent digestibility coefficient (ADC) was 92.4% for dry matter, 91.4% for crude protein, and 94.2% for crude lipid in rainbow trout. In the second trial, six diets were formulated to replace FO at 0% (CON), 20% (T20), 40% (T40), 60% (T60), 80% (T80), and 100% (T100) with SC in the rainbow trout (3.0 ± 0.4 g, mean ± SD) diet. After eight weeks' feeding trial, weight gain (WG), specific growth rate (SGR), and feed efficiency (FE) of fish fed the T20 diet were significantly higher than those of fish fed other diets (p < 0.05). However, there were no significant differences in these parameters among those of fish fed CON, T40, T60, and T80 diets. Lysozyme activity of fish fed the T20 diet was significantly higher than those of fish fed other experimental diets (p < 0.05). After 10 days of disease challenge testing with pathogenic bacteria (Lactococcus garvieae 1 × 108 CFU/mL), the cumulative survival rate of fish fed the T20 diet was significantly higher than those of fish fed the CON, T80, and T100 diets. Therefore, these results suggest dietary microalgae SC is well-digested and could replace up to 80% of fish oil in the diet of rainbow trout without negative effects on growth and immune responses.

Synergistic effects of dietary supplementation of Bacillus subtilis WB60 and mannanoligosaccharide (MOS) on growth performance, immunity and disease resistance in Japanese eel, Anguilla japonica.

This study evaluated the synergistic effects of dietary Bacillus subtilis WB60 and mannanoligosaccharide (MOS) in juvenile Japanese eel, Anguilla japonica. Seven treatment diets were formulated to contain three different levels of B. subtilis (0.0, 0.5, and 1.0 × 107 CFU/g diet denoted as BS0, BS0.5, and BS1, respectively) with two MOS levels (0 and 5 g/kg diet denoted as M0 and M5, respectively), and one diet with oxytetracycline (OTC) at 5 g/kg diet. Each diet (BS0M0 (CON), BS0M5, BS0.5M0, BS0.5M5, BS1M0, BS1M5, and OTC) was fed to triplicate groups of 20 fish averaging 9.00 ±â€¯0.11 g (mean ±â€¯SD) for eight weeks. Average weight gain, feed efficiency, specific growth rate and protein efficiency ratio of fish fed the BS0.5M5 and BS1M5 diets were significantly higher than those of fish fed CON, BS0.5M0 and OTC diets (P < 0.05). Significant increases in the nonspecific enzymatic activities (e.g., lysozyme and myeloperoxidase) were detected from fish fed the BS0.5M5, BS1M5, and OTC diets compared to the CON, BS0.5M0, and BS0M5 diets (P < 0.05). Whereas, immunoglobulin M expressions were recorded significantly higher for fish fed the BS0.5M5 and BS1M5 diets compared to those of fish fed the other diets (P < 0.05). Also, heat shock protein 70 mRNA levels of fish fed BS0.5M5 and BS1M5 diets were significantly higher than those of fish fed the CON diet (P < 0.05). Histological observations of the intestinal morphology showed healthier gut for fish fed BS0.5M5 and BS1M5 diets than those fed CON, BS0M5, and OTC diets. Additionally, resistance to bacterial challenge with Vibrio anguillarum was recorded significantly lower for fish fed the CON diet than those fed other diets (P > 0.05). Therefore, the results for growth performance, non-specific immune responses, intestinal morphology, and disease resistance demonstrated that supplementation of B. subtilis at 0.5 × 107 CFU/g diet and mannanoligosaccharide at 5 g/kg diet could have beneficial synergistic effects in Japanese eel. The isolated probiotic from eel and the selected prebiotic could lead to the development of a specific and potential synbiotic in Japanese eel aquaculture.

Comparative evaluation of dietary probiotics Bacillus subtilis WB60 and Lactobacillus plantarum KCTC3928 on the growth performance, immunological parameters, gut morphology and disease resistance in Japanese eel, Anguilla japonica.

The current experiment was conducted to evaluate and compare the efficacy of two different probiotics Bacillus subtilis WB60 and Lactobacillus plantarum KCTC3928 in diet of Japanese eel, Anguilla japonica. Seven experimental diets were formulated to contain no probiotics (CON), three graded levels of B. subtilis at 106 (BS1), 107 (BS2), 108 (BS3) and L. plantarum at 106 (LP1), 107 (LP2), 108 (LP3) CFU/g diet. Twenty fish averaging 8.29 ± 0.06 g were distributed in to 21 aquaria and were randomly assigned to one of the experimental diets in triplicate groups. Average weight gain (WG), feed efficiency (FE), and protein efficiency ratio (PER) of fish fed B. subtilis at 107 (BS2) and 108 (BS3) CFU/g diet were significantly higher than those of fish fed other experimental diets (P < 0.05). Nonspecific enzymatic activities including lysozyme, superoxide dismutase (SOD), myeloperoxidase (MPO) from fish fed B. subtilis at 107 (BS2) and 108 (BS3) CFU/g diet were detected to be significantly higher than that from fish fed CON diet (P < 0.05). Whereas, level of intestine glyceraldehyde-3-phosphate dehydrogenase (GAPDH), heat shock protein 70, 90 (HSP70, 90) and immunoglobulin (IgM) from fish fed B. subtilis at 107 and 108 CFU/g diet were recorded to be significantly higher than those of fish fed other experimental diets (P < 0.05). Parameters for intestinal morphology and histology suggested a healthier gut for the fish fed108 (BS3) CFU/g diet as compared to other treatment groups. Whereas, results from the disease challenge test with bacteria Vibrio angulillarum showed significantly lower survival rate for fish fed CON diet than those of fish fed other experimental diets. Therefore, these results indicated that oral supplement of B. subtilis at 108 (BS3) CFU/g diet could be a more effective source of probiotic compared to L. plantarum in Japanese eel.

Evaluation of Optimum Dietary Threonine Requirement by Plasma Free Threonine and Ammonia Concentrations in Surgically Modified Rainbow Trout, Oncorhynchus mykiss.

This study was carried out to evaluate the dietary threonine requirement by measuring the plasma free threonine and ammonia concentrations in rainbow trout, Oncorhynchus mykiss after dorsal aorta cannulation. A total of 70 fish (average initial weight 506±8.2 g) were randomly distributed into each of the 14 net cages (5 fish/cage). After 48 hours (h) of feed deprivation, each group was intubated at 1% body weight with one of the seven L-amino acid based diets containing graded levels of threonine (0.42%, 0.72%, 0.92%, 1.12%, 1.32%, 1.52%, or 1.82% of diet, dry matter basis). Blood samples were taken at 0, 5, and 24 h after intubation. Post-prandial plasma free threonine concentrations (PPthr) of fish 5 h after intubation with diets containing 1.32% or more threonine were significantly higher than those of fish intubated with diets containing 1.12% or less threonine (p<0.05). Post-absorptive free threonine concentrations (PAthr) after 24 h of intubation of the fish with diets containing 0.92% or more threonine were significantly higher than those of fish intubated with diets containing 0.72% or less threonine. Post-prandial plasma ammonia concentrations (PPA, 5 h after intubation) were not significantly different among fish intubated with diets containing 1.12% or less threonine, except the PPA of fish intubated with diet containing 0.42% threonine. Broken-line model analyses of PPthr, PAthr, and PPA indicated that the dietary threonine requirement of rainbow trout should be between 0.95% (2.71) and 1.07% (3.06) of diet (% of dietary protein on a dry matter basis).

Evaluation of the Efficacy of Fermented By-product of Mushroom, Pleurotus ostreatus, as a Fish Meal Replacer in Juvenile Amur Catfish, Silurus asotus: Effects on Growth, Serological Characteristics and Immune Responses.

The present experiment was conducted to evaluate the efficacy of dietary fermented by-product of mushroom, Pleurotus ostreatus, (FBPM) as a fish meal (FM) replacer in juvenile Amur catfish, Silurus asotus. A total number of 225 fish averaging 5.7±0.1 g (mean±standard deviation) were fed one of the five experimental diets formulated to replace FM with FBPM at 0%, 5%, 10%, 20%, and 30% (FBPM0, FBPM5, FBPM10, FBPM20, and FBPM30, respectively). At the end of eight weeks of the experiment, average weight gain (WG) of fish fed FBPM0 or FBPM5 were significantly higher than those of fish fed FBPM20 or FBPM30 diets (p<0.05). However, there was no significant differences in WG among the fish fed FBPM0, FBPM5 or FBPM10, and between fish fed FBPM10 or FBPM20, and also between those fed FBPM20 or FBPM30 diets. Lysozyme activity of fish fed FBPM0 or FBPM5 were significantly higher than those of fish fed FBPM10, FBPM20 or FBPM30 diets (p<0.05). The chemiluminescent response of fish fed FBPM5 was significantly higher than those of fish fed FBPM0, FBPM20 or FBPM30 diets (p<0.05). Broken line regression analysis of WG suggested that the maximal dietary inclusion level for FBPM as a FM replacer could be 6.3% without any adverse effects on whole body composition and on serological characteristics. Therefore, these results may indicate that the maximal dietary inclusion level of FBPM as a FM replacer could be 6.3% in juvenile Amur catfish.