Intestinal microbiota of white shrimp,Litopenaeus vannamei,fed diets containing Bacillus subtilis E20‐fermented soybean meal (FSBM) or an antimicrobial peptide derived from B. subtilis E20‐FSBM

The purpose of the present study is to profile the microfloral structure in the intestines of the white shrimp, Litopenaeus vannamei, after being fed a control diet, in comparison with the diets containing Bacillus subtilis E20‐fermented soybean meal (FSBM) or an antimicrobial peptide (AMP) isolated from B. subtilis E20‐FSBM (diets designated FSBMD and AMPD, respectively) for 60 days. Metagenomic data showed detection of eight phyla, 20 classes, 40 orders, 68 families and 96 genera. Despite no statistically significant difference, an evaluation of microbial diversity recorded higher species richness, Shannon–Weaver diversity index and evenness in the AMPD and FSBMD groups, compared to the control diet group. Venn diagrams showed that 58 of the operational taxonomic units (OTUs) were shared among the control, FSBMD and AMPD groups, but additional beneficial microbes were only found in the FSBMD and AMPD groups. In addition to the augmentation of beneficial bacteria in the FSBMD and AMPD groups, the abundance of potential pathogens, Vibrio and Flavobacterium, was lower in the gut of shrimp fed the FSBMD and AMPD. The results clearly suggest that the FSBMD and AMPD have the capability to change the microfloral structure of shrimp intestines and could be used for disease prevention in shrimp aquaculture.     

Administration of probiotic Bacillus licheniformis induces growth,immune and antioxidant enzyme activities,gut microbiota assembly and resistance to Vibrio parahaemolyticus in Litopenaeus vannamei

This study aimed to elucidate the effects of dietary Bacillus licheniformis ATCC 11946 (BL) on the growth, immune and antioxidant activities, intestinal morphology and microbiota, and susceptibility to Vibrio parahaemolyticus (VP_AHPND) in juvenile pacific white shrimp, Litopenaeus vannamei (LV). Juvenile LV (initial weight = 0.63 ± 0.001) were fed diets containing varying BL concentrations (0 (BL0), 10⁶ (BL1), 10⁷ (BL2) and 10⁸ (BL3) CFU/g feed) for 8 weeks. Growth performance, immune and antioxidant enzyme activities, and intestinal morphology significantly improved in the probiotic‐treated groups than the untreated. Regardless of the treatment group, the two dominant phyla were Proteobacteria and Bacteroidetes, whereas the two dominant genera were Ruegeria and Vibrio. Increasing inclusion of probiotics in diets led to significant increase in beneficial bacterial genera (Ruegeria, and Pseudoalteromonas) and a significant decrease in some known opportunistic pathogens (Vibrio, Tenacibaculum, Photobacterium, Kangiella and Spongiimonas) with the BL3 group witnessing the best. A 7‐day challenge study with VP_AHPND showed significantly high protection in the probiotic‐treated groups, with the BL3, BL2 and BL1 obtaining 51%, 34% and 22% relative percentage survival, respectively. In conclusion, BL at 10⁸ CFU/g feed should be used to help in shrimp production since it attained the most significant improvement.     

Effects of fish origin probiotics on growth performance,immune response and intestinal health of shrimp (Litopenaeus vannamei) fed diets with fish meal partially replaced by soybean meal

This study was conducted to evaluate the application of four fish origin probiotics to relieve the side effects induced by SBM in shrimp. Shrimps were fed with high fish meal diet (C0, positive control containing 500 g/kg FM and 0 g/kg SBM), low fish meal diet (C, control containing 250 g/kg FM and 340 g/kg SBM) or the control diet (C) supplemented, respectively, with 1.0 × 10⁸ CFU/g Bacillus pumilus SE5 (T1), Psychrobacter sp. SE6 (T2), Enterococcus faecium MM4 (T3) or Bacillus claussi DE5 (T4). After 6 weeks of feeding, the growth performance, hepatopancreatic digestive enzymes, intestinal histological structure and immune parameters were determined. The results showed that compared with high FM group (C0), significantly lower WGR and SGR as well as higher FCR were observed in the low FM group (C) (p < .05). Compared with the control, significantly lower FCR was observed in treatment T1 (p < .05), but not in treatments T2–T4. Protease, amylase and lipase activities in all the treatments (T1‐T4) were significantly higher than the control (p < .05). GOT and GPT activities in the control were significantly lower than the C0 (p < .05), while higher MDA level was recorded in the control (p < .05). Meanwhile, higher GPT activities were observed in treatments T1‐T4 compared with the control (p < .05). Compared with the C0, lower SOD, ACP and AKP activities were observed in the control. Significantly improved SOD and AKP activities were observed in all probiotic feeding groups compared with the control (p < .05). The control diet led to significant reduction of intestinal wall thickness and villus height compared with the C0 (p < .05), while significantly higher intestinal wall thickness and villus height were exhibited in all the probiotic groups compared with the control (p < .05), except intestinal wall thickness in treatment T3. Thus, these results demonstrated that the four fish origin probiotic strains could relieve the side effects induced by high level of SBM in shrimp, while probiotic B. pumilus SE5 showed the best performance.     

Beneficial effects of dietary exogenous protease on the growth,intestinal health and immunity of GIFT (Oreochromis niloticus) fed plant‐based diets

This study investigated the effects of dietary exogenous protease on the growth performance, intestinal health, immune parameters and disease resistance of genetically improved farmed tilapia (GIFT, Oreochromis niloticus). Five test diets with commercial protease at the levels of 0, 1.38, 2.76, 5.52 and 11.04 U/g (named PE0, PE1, PE2, PE5 and PE11, respectively) were administered to triplicate tanks with 30 fish for 60 days, and then, the fish were challenged with Streptococcus agalactiae for 14 days. The results indicated that weight gain increased as exogenous protease increased from 0 to 5.52 U protease/g diet and then decreased significantly (p < .05) with a further increase in exogenous protease supplementation (p < .05). The height of the villi in the proximal intestine and distal intestine, the width of the villi in three segments of the intestine, and the thickness of the muscle layer in the proximal intestine and mid‐intestine (p < .05) were increased in the fish fed the PE5 diet. Immune and antioxidant indices (except malondialdehyde), and survival after challenged with S. agalactiae were higher in fish fed PE5 diets than in those fed other diets (p < .05). In conclusion, 5.52 U/g protease supplementation in a plant‐based diet could promote the growth performance, intestinal physical barrier function, innate immunity and S. agalactiae resistance of GIFT.     

Effects of bioflocs with different C/N ratios on growth,immunological parameters,antioxidants and culture water quality in Opsariichthys kaopingensis Dybowski

Biofloc technology (BFT) is a new ecological aquaculture technology that is intended not only to eradicate pollutants and elevate feed utilization but also to enhance immunity and antioxidant activity in aquatic animals. A 28‐day feeding trial was implemented to evaluate the effects of promoted bioflocs on the water quality, growth performance, immunological parameters and antioxidant status of Opsariichthys kaopingensis Dybowski juveniles in low exchange culture tanks. Three hundred sixty healthy O. kaopingensis juveniles (7.1 ± 0.02 g) were irregularly distributed among 12 tanks. Four C:N ratios in triplicate tanks were tried: C/N = 10.8:1 with a commercial diet (control), C/N = 15:1, C/N = 20:1 and C/N = 25:1. Anhydrous glucose (99.97%) was added to the commercial diet to maintain elevated carbon ratio in the three treatments. Uninterrupted 24‐hr aeration was supplied during the test by using an air‐stone connected to an air pump to develop the bioflocs in the water column. The results indicated that BFT significantly reduced total ammonia nitrogen (TAN), nitrite (NO2^?‐N), total phosphorus (TP) concentrations and water transparency (Secchi depth) in C/N 15, C/N 20 and C/N 25 (p < .05), whereas the chemical oxygen demand (COD) and nitrate nitrogen (NO3^?‐N) concentrations gradually increased over time. In addition, weight gain, specific growth rate (SGR), survival and protein efficiency ratio (PER) were significantly elevated in C/N 20, while food conversion rate (FCR) was considerably decreased (p < .05) in C/N 20, compared with the control. Significant increases in lysozyme (LSZ), glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT) and alkaline phosphatase (AKP) activities of O. kaopingensis were found in the C/N 20 group after a 28‐day feeding trial (p < .05). Comparing the antioxidant capacity of O. kaopingensis in gills, brains, kidneys, hepatopancreas, intestines and serum of juveniles from the four experimental groups, the activity of superoxide dismutase (SOD), total antioxidant capacity (T‐AOC), catalase (CAT) activity and glutathione peroxidase (GSH‐PX) activity of juveniles in the C/N 20 group were significant higher (p < .05), and the content of malondialdehyde (MDA) was considerably lower, than in the control. Overall, these findings suggest that BFT not only can enhance O. kaopingensis growth performance and strengthen antioxidant status but also can upgrade immune response, ensuring the sustainable development of aquaculture.     

Partial replacement of fishmeal with Clostridium autoethanogenum single‐cell protein in the diet for juvenile black sea bream (Acanthopagrus schlegelii)

Clostridium autoethanogenum protein (CAP) is a single‐cell protein derived from ethanol production and may have potential to become a substitute for fishmeal in aquafeeds. A 70‐day feeding trial was conducted with black sea bream (Acanthopagrus schlegelii) (mean initial weight 6.03 ± 0.09 g) to evaluate partial replacement of fishmeal with CAP in diets. Seven isonitrogenous and isoenergetic diets were formulated with graded levels of CAP (0, 4.85, 9.70, 14.55, 19.40, 38.80 and 58.20%) to replace fishmeal. The inclusion of CAP at all dietary levels tested did not significantly affect the growth performance (p > .05). Fish fed the CAP58.20% diet showed a significantly lower feeding rate, with significantly higher protein efficiency ratio and feed efficiency ratio compared with fish fed the other diets (p < .05). No statistical differences were found in dorsal muscle and whole‐body compositions. Total superoxide dismutase in serum of fish fed CAP58.20% diet was significantly lower compared with that of the control. Malondialdehyde, catalase, total antioxidant capacity and digestive enzyme activities revealed no significant differences among dietary treatments. Phosphorus retention efficiency significantly increased, and phosphorus discharge showed a downward trend with increasing CAP inclusion levels. In conclusion, the results indicated that CAP is a safe and effective alternative protein source, which can replace fishmeal in the diet of black sea bream up to 58.20%, without adverse effects on growth performance, antioxidation and digestive enzyme activity. This study has shown the potential of converting industrial waste into a high protein feed ingredient for aquafeeds.     

Effects of salinity level on the survival,growth, feed utilization,carcass composition,haematological and serum biochemical changes of juvenile Meagre (Argyrosomus regius) (Asso, 1801) grown in ground saltwater

A study was performed to examine the effects of salinity on water quality, fish performance, carcass composition and haemato‐biochemical parameters in juvenile meagre, Argyrosomus regius. Fish (5.0 g) were stocked in fibreglass tanks at four salinity levels: 8‰, 16‰, 24‰ and 32‰, and fed a pelleted diet (47/17 protein/lipid) for 56 days. Results indicated that the growth, feed utilization, carcass composition and haemato‐biochemical parameters of meagre gradually improved with the increase in salinity up to 24‰ and then significantly (p ≤ .05) decreased at 32‰. The survival per cent showed a significant decrease when A. regius exposed to 8‰ salinity. An improvement with 32%, 47% and 34.1% of FCR, protein productive value and energy utilization was detected at 24‰ compared with 8‰ salinity respectively. The highest content of protein and the lowest of lipids were recorded in fish carcass at 24‰ compared with the opposite trend at 8‰ salinity. The 24‰ salinity treatment exhibited the highest value of haemoglobin (4.9 g/dl) and the lowest ratio (0.73) of albumin/globulin. The serum total protein, albumin and globulin were significantly higher at 24‰ and 32‰ salinity than those at 8‰ and 16‰ salinity groups. These findings indicate that 24‰ salinity level might be the best for meagre.     

Effects of glycinin on growth performance,immunity and antioxidant capacity in juvenile golden crucian carp,Cyprinus carpio × Carassius auratus

This study was designed to examine the effects of glycinin on growth, digestive ability, immune responses, antioxidant capacity and gene expression levels of golden crucian carp. Golden crucian carp were fed diets containing glycinin at 0, 30, 60, 90 and 120 g/kg, respectively, for 8 weeks. Body weight, weight gain percentage, specific growth rate and feed efficiency ratio were negatively related to the content of glycinin in diet. Activities of protease, acid phosphatase, alkaline phosphatase, lysozyme in hepatopancreas, and activities of catalase, glutathione peroxidase, superoxide dismutase, and total antioxidant capacity in the proximal intestine, mid intestine, distal intestine and hepatopancreas were negatively related to the content of glycinin in diet, whereas malondialdehyde in proximal intestine, mid intestine, distal intestine and hepatopancreas increased directly with the content of glycinin in diet. Furthermore, the relative expressions of TNF‐α and IL‐1β in proximal intestine, mid intestine and distal intestine increased directly with the content of glycinin in diet, whereas the relative expressions of TNF‐α and IL‐1β in hepatopancreas were negatively related to the content of glycinin in diets. The relative expressions of IL‐10 in proximal intestine, mid intestine, distal intestine and hepatopancreas all were negatively related to the content of glycinin in diets. In conclusion, reductions in growth, immunity and antioxidant capacity, intestine inflammation with dysfunction of digestive system occurred in golden crucian carp that fed a diet containing glycinin at 30 g/kg or higher after 8 weeks.     

Non‐spherical pearl layers in the Polynesian ‘black‐lipped’ Pinctada margaritifera: The non‐nacreous deposits compared to microstructure of the shell growing edge

Cultivated pearls frequently exhibit morphological irregularities making obvious that mineral deposition was irregularly distributed onto nucleus surface. Taking advantage of experimental cultivations with short durations (from 10 days to few months), these irregular deposits predating occurrence of the nacre were investigated in Polynesian pearls by biochemical characterizations and a series of physical methods. Diversity in the resulting data suggests that various in‐depth alterations of the biomineralization mechanism may have occurred during the grafting process, leading to diversity in the biochemical pathways to nacreous deposition. This allows a precise discussion of current views about pearl formation. The “reversed shell theory” is formally disproved through point to point comparison with development of the shell growing edge. Similarity of pearl formation with “regeneration” or “shell repair” is also discussed, emphasizing the differences between these concepts.