Dietary Clostridium autoethanogenum protein has dose-dependent influence on the gut microbiota, immunity, inflammation and disease resistance of abalone Haliotis discus hannai.

Clostridium autoethanogenum protein (CAP) is an eco-friendly protein source and has great application potential in aquafeeds. The present study aimed to investigate the effects of dietary CAP inclusion on the anti-oxidation, immunity, inflammation, disease resistance and gut microbiota of abalone Haliotis discus hannai after a 110-day feeding trial. Three isonitrogenous and isolipidic diets were formulated by adding 0% (control), 4.10% (CAP4.10) and 16.25% (CAP16.25) of CAP, respectively. A total of 540 abalones with an initial mean body weight of 22.05 ± 0.19 g were randomly distributed in three groups with three replicates per group and 60 abalones per replicate. Results showed that the activities of superoxide dismutase and glutathione peroxidase in the cell-free hemolymph (CFH) were significantly decreased and the content of malondialdehyde in CFH was significantly increased in the CAP16.25 group. The diet with 4.1% of CAP significantly increased the activities of lysozyme and acid phosphatase in CFH. The expressions of pro-inflammatory genes such as tumor necrosis factor-α (tnf-α), nuclear factor-κb (nf-κb) and toll-like receptor 4 (tlr4) in digestive gland were downregulated, and the expressions of anti-inflammatory genes such as ß-defensin and mytimacin 6 in digestive gland were upregulated in the CAP4.10 group. Dietary CAP inclusion significantly decreased the cumulative mortality of abalone after the challenge test with Vibrio parahaemolyticus for 7 days. Dietary CAP inclusion changed the composition of gut microbiota of abalone. Besides, the balance of the ecological interaction network of bacterial genera in the intestine of abalone was enhanced by dietary CAP. The association analysis showed that two bacterial genera Ruegeria and Bacteroides were closely correlated with the inflammatory genes. In conclusion, the 4.10% of dietary CAP enhanced the immunity and disease resistance as well as inhibited the inflammation of abalone. The 16.25% of dietary CAP decreased the anti-oxidative capacity of abalone. The structure of the gut microbiota of abalone changed with dietary CAP levels.

Beneficial microbes to suppress Vibrio and improve the culture performance of copepod Tigriopus japonicus Mori.

The use of beneficial microbes, i.e., probiotics, to reduce pathogens and promote the performance of the target species is an important management strategy in mariculture. This study aimed to investigate the potential of four microbes, Debaryomyces hansenii, Ruegeria mobilis, Lactobacillus plantarum, and Bacillus subtilis, to suppress Vibrio and increase survival, population growth and digestive enzyme activity (protease, lipase, and amylase) in the harpacticoid copepod, Tigriopus japonicus. Copepod, T. japonicus stock culture with an initial mean density of 50 individual/mL (25 adult male and 25 adult female) was distributed into five treatments (i.e., four experimental and a control, each with four replicates; repeated twice) using 20 beakers (100 mL capacity each). The copepods were fed a mixture of the dinoflagellate Alexandrium tamarense and the diatom Phyaeodactylum tricornutum (3 × 104 cells/mL-1). Each microbe's concentration was adjusted at 108 CFU/mL-1 and applied to the culture condition. D. hansenii, L. plantarum, and B. subtilis all improved the copepods' survival and population growth, likely by including a higher lipase activity (P < 0.05). In contrast, using R. mobilis did not improve the copepod's culture performance compared to control. B. subtilis was the most effective in decreasing the copepod's external and internal Vibrio loading. The probiotic concentrations in the copepod decreased within days during starvation, suggesting that routine re-application of the probiotics would be needed to sustain the microbial populations and the benefits they provide. Our results demonstrated that D. hansenii and B. subtilis are promising probiotics for mass copepod culture as live food for mariculture purposes.

The Use of Selected Bacteria and Yeasts to Control Vibrio spp. in Live Food.

Vibrio species are a significant causative of mass mortality in mariculture worldwide, which can quickly accumulate in live food and transmit into the larval gut. With restrictions on the use of antibiotics in aquaculture, finding a proper solution to reduce the risk of Vibriosis is vital. This study aimed to evaluate the susceptibility of Vibrio harveyi, V. campbellii, V. anguillarum, and V. parahaemolyticus to twenty-six bacterial and yeast strains and use the beneficial ones to enrich live food (Branchiopod, Artemia franciscana, rotifer, Brachionus plicatilis and copepod, Tigriopus japonicus). Thus, a modified disk diffusion method was applied. After a susceptibility assay, the bacteria and yeast beneficial in suppressing the Vibrio species were labeled by fluorescent stain and used to measure the accumulation potential in different live foods. Also, the beneficial bacteria and yeast were used to enrich live foods, and then the count of loaded Vibrio was estimated after 5, 10, 15, and 20 hours by the serial dilution method. From the total bacteria and yeast strains that were used, Candida parapsilosis, Pseudoalteromonas flavipulchra, Lactobacillus sakei, Bacillus natto, and B. amyloliquefaciens inhibited all four Vibrio species. The results of microbial labeling showed that L. sakei in Artemia, C. parapsilosis in rotifers, and V. harveyi in copepods had the highest accumulation rate. The results of the estimation of loaded Vibrio in different live foods also showed that the use of beneficial bacteria and yeast each significantly reduced the count of Vibrio. Application of bacteria and yeast to suppress pathogenic Vibrio maybe a sustainable method for preventing this pathogen from harmfully invading aquaculture and may also aid in reducing the chances of antibiotic resistance in pathogenic Vibrio.

The Use of Two Bifidobacterium Strains Enhanced Growth Performance and Nutrient Utilization of Rainbow Trout (Oncorhynchus mykiss) Fry.

This study aims to evaluate the effects of Bifidobacterium animalis PTCC-1631 and Bifidobacterium lactis PTCC-1736 on growth, nutrient digestibility, carcass composition, and intestinal microbiota of Oncorhynchus mykiss fry, which had initial body weights of 0.58 ± 0.19 g. Four iso-nitrogenous diets with probiotics, namely T1 (1 × 107 CFU g-1), T2 (2 × 107 CFU g-1), T3 (3 × 107 CFU g-1), and control without probiotics were obtained. Total 480 fish were randomly divided in four treatments (three experimental treatments and a control), each with 4 replicates including 16 tanks each filled with 15 L-1 of water (water exchange rate of 2 L min-1 and aerated with air stone) with a density of 2 fry L-1. The water mean temperature was 17.66 ± 1.33 °C, and the mean pH value was 7.63 ± 0.08. After eight weeks of the experiment T1 that fed with a diet contained the lowest bacterial concentration showed higher growth, nutrient utilization, digestibility, and lower feed conversion ratio (P < 0.05) in comparison with T2, T3, and the control. Also, the highest count of lactobacillus bacteria was observed in T1 that was fed with a diet containing the lowest probiotic. The lowest concentration of Bifidobacterium strains in this study induced better growth and increased digestion and nutrient utilization in trout fry.