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Abstract: In recent years, more and more attentions have been paid to the development and application of probiotics in aquaculture, and viable probiotics have been extensively studied, while rare information was available about inactivated probiotics in aquaculture. Therefore, in this study, a feeding trial was designed to investigate the effect of dietary supplementation of heat‐inactivated probiotic Bacillus clausii DE5 on growth performance, immune response and key immune genes expression in head kidney and intestine in grouper Epinephelus coioides. Fish were fed for 60 days with control diet (C) and two experimental diets containing 1.0 × 108 CFU/g live (T1) and heat‐inactivated (T2) B. clausii DE5, respectively. The probiotic treatments did not affect the final weight, weight gain (WG) and specific growth rate (SGR) of E. coioides at days 30 and 60 (p > .05), while both heat‐inactivated and live B. clausii DE5 significantly decreased the feed intake and feed conversion ratio (FCR) at day 60 (p < .05). Serum lysozyme activity and complement C3 level in the two probiotic treatments were significantly higher than those in the control (p < .05). The lysozyme activity and complement C3 level at day 60 were significantly higher than those at day 30 (p < .05), while no significant interaction effect between diet and administration date was observed. Moreover, the heat‐inactivated B. clausii DE5 significantly improved the expression of TLR5, pro‐inflammatory cytokines (IL‐8 and IL‐1β) and TGF‐β1 in head kidney and intestine (p < .05), while the live probiotic did not show any significant effect on the expression of these key immune‐related genes in head kidney and intestine. These results indicate that dietary supplementation of heat‐inactivated B. clausii DE5 effectively improved feed utilization and both the local and systemic immune responses of E. coioides. [ABSTRACT FROM AUTHOR]

To evaluate the possible dietary application of live and heat-inactivated probiotic Bacillus pumilus SE5 in grouper Epinephelus coioides, juveniles (14.6 ± 0.2 g) were fed either a basal control diet (without probiotic) or the basal diet supplemented with 1.0 × 108 CFU g−1 live (T1) and heat-inactivated B. pumilus SE5 (T2). The heat-inactivated probiotic significantly improved the final weight, weight gain ( WG) and specific growth rate ( SGR) at day 60 and significantly decreased the feed conversion ratio ( FCR) at day 30 and 60, while the viable probiotic significantly decreased the FCR at day 60 ( P < 0.05). Phagocytic activity, serum complement C3 and IgM levels as well as SOD activity elevated significantly in fish fed the heat-inactivated probiotic for 60 days ( P < 0.05). Furthermore, the heat-inactivated probiotic remarkably up-regulated expression of TLR2 and pro-inflammatory cytokines ( IL-8 and IL-1β) in head kidney ( P < 0.05), but the viable probiotic failed to do so. These results indicated that heat-inactivated B. pumilus SE5 can effectively improve the growth performance and immune responses of E. coioides. [ABSTRACT FROM AUTHOR]

The effect of dietary administration of Lactococcus lactis MM1 and Enterococcus faecium MM4, two strains of lactic acid bacteria isolated from the gut of juvenile grouper Epinephelus coioides, on the growth, feed utilization, digestive enzymes and immune response of juvenile E. coioides was evaluated. The fish were fed for 60 days with three different diets: control diet (without probiotic), diet P1 supplemented with 1.0 × 108 CFU g−1 L. lactis or diet P2 with 1.0 × 108 CFU g−1 E. faecium. Although growth and feed intake decreased slightly, significant improvement of feed utilization was observed in fish fed diet P1 and diet P2 for 60 days. The hepatopancreatic protease activities in the two probiotic treatments were significantly higher than that in the control. The phagocytic activity, serum superoxide dismutase activity and complement component 4 (C4) and IgM levels were not significantly affected by diet P1 and diet P2. Serum complement component 3 (C3) levels in the two treatments were significantly higher than that in the control. Significant increased serum lysozyme activity was observed in fish fed the diet P1, while significant decreased lysozyme activity was observed in fish fed the diet P2 compared with the fish fed the control diet. There was a significant interaction effect between diet and administration date for the complement C4. The study demonstrates potential for L. lactis MM1 and E. faecium MM4 to improve the feed utilization and modulate the immune function in E. coioides. [ABSTRACT FROM AUTHOR]

A 60-day feeding trial was conducted to assess the effect of incorporating dietary Psychrobacter sp., one predominant bacterium with antagonistic activity in the gut of fast-growing grouper Epinephelus coioides, on the growth, feed utilization, digestive enzymes and immune responses of E. coioides at regular monthly intervals. The fish were fed with control diet (without probiont) or probiotic diet supplemented with 1.0 × 10 CFU g Psychrobacter sp. No significant improvement in weight gain (WG) and specific growth ratio (SGR) were observed in the probiotic group, but a significant improvement in feed conversion ratio (FCR) was observed in fish fed the probiotic diet for 60 days. Although not statistically significant, enhanced hepatopancreatic protease and lipase activities and intestinal amylase activity were observed in probiotic group at day 60. A slightly increase in phagocytic activity and phagocytic index was observed in fish fed probiotic diet for 60 days. Serum lysozyme activity showed no significant differences between the probiotic group and the control group during the whole experiment period, while an increasing trend of superoxide dismutase (SOD) activity in the probiotic group was observed. Serum complement component 3 (C3) level of the probiotic group was higher than that of control in the whole feeding period, while complement component 4 (C4) level of the probiotic group was significantly higher than that of the control at day 60. The results of the current study firstly demonstrated potential for Psychrobacter sp. as a novel probiont to improve feed utilization, digestive enzymes and innate immunity in E. coioides. [ABSTRACT FROM AUTHOR]

Aims: To evaluate the diversity of dominant autochthonous microbiota along the digestive tract of juvenile Epinephelus coioides following the dietary administration of probiotic Bacillus pumilus for 60 days. Methods and Results: Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) with subsequently sequencing analysis was used to assess the gut microbiota . Generally similar DGGE patterns were observed in the foregut, midgut and hindgut of E. coioides, while the similarity dendrogram clearly revealed three different clusters depending on the three compartments of the GI tract. Dietary administration of B. pumilus stimulated its colonization in each compartment of the digestive tract. Samples collected from the probiotic group and the control group showed similar DGGE patterns, and no significant difference in the total number of bands and the Shannon index were detected between the probiotic group and the control group, suggested that B. pumilus exert no significant effect on the gut microbiota. However, various potentially beneficial bacteria, such as uncultured Bacillus sp. clone QJNY94-like, Nitratireductor sp. YCSC5-like, Methylobacterium hispanicum-like and Microbacterium sp. YACS1-like bacteria were stimulated by probiotic B. pumilus, while the potential harmful Staphylococcus saprophyticus-like bacterium was depressed. Conclusions: Autochthonous gut microbiota of E. coioides was modulated to some degree, not significant, by probiotic B. pumilus, various potentially beneficial bacteria were selectively stimulated, while one potential harmful species was depressed. Significance and Impact of the Study: This work represents the first report that dietary administration of probiotic B. pumilus modulated the gut microbiota of E. coioides. These findings broaden our understanding of probiotic effects at the gut level, which is helpful in understanding the mechanisms that underpin host benefits. [ABSTRACT FROM AUTHOR]

Abstract: The relationship(s) between nodavirus infection and myostatin expression in the skeletal muscle tissue of grouper is unclear. To investigate, the grouper (Epinephelus coioides) myostatin gene was cloned and cDNA was utilized to examine the expression of the gene in skeletal muscle and serum of healthy (uninfected) grouper and fish naturally infected with nodavirus. The myostatin gene comprises three exons and two introns and is transcribed as a 2778-bp mRNA length that encodes a 376-aa precursor protein. All exon–intron boundaries conformed to the consensus sequences. Alignment of the upstream sequences indicated that the grouper myostatin promoter has been highly conserved during evolution. Sequence analyses of 1936 bp of the upstream region revealed ten E-box motifs. The protein was consistent with the predicted molecular weight (approximately 42 kDa) of the unprocessed monomeric precursor protein and the processed myostatin form of the protein secreted into the plasma. Transient transfection studies revealed that the activity of the myostatin promoter decreased in a subset of viral titers. Grouper naturally infected with nodavirus displayed downregulation of the myostatin protein. [Copyright &y& Elsevier]

The histological architecture of major immune organs in the Malabar grouperEpinephelus malabaricuswas investigated. The novel characteristics such as melanomacrophage centres(MMCs) appeared in the thymus and lymphopoietic tissue formed as foci in the head kidney. Leukocyte distribution in organs was identified by enzyme histochemistry.β-glucuronidase(BG) reactive cells in the cortex region of the thymus were botryoidally aggregated. Both acid phosphatase(AcP) and BG reactive cells concentrated within the specialized lymphopoietic foci in the head kidney, suggesting that the foci might be functional. Primitive histological characters of immunity were observed in the spleen. Although leukocyte aggregation was demonstrated in the spleen, additional enzyme histochemistry indicated that the aggregate might not be the equivalent of white pulp found in other vertebrates. The histological evidence did not support intestinal involvement in the immune system: there was no demonstrable gut associated lymphoid tissue. The limited distribution in the cortex and medulla boundary and the condensed format of the lymphocytes suggests a functional role for the MMC in the thymus ofE. malabaricus. The MMC appearance in the thymus of a teleost was unusual. [ABSTRACT FROM AUTHOR]