Partial replacement of fish oil with terrestrial lipid blend and effects on growth performance,body composition,immune parameter and growth‐related genes in yellow drum (Nibea albiflora)

The imbalance of fish oil (FO) supply and demand has motivated efforts to identify an alternative for aqua feed. An 8‐week feeding trial was conducted to evaluate the effects of dietary fish oil partial replacement with mixed oil from linseed and lard (1:1) on growth, body composition and immunity when the N3 long‐chain polyunsaturated fatty acid (N3 LC‐PUFAs) requirement is met for Nibea albiflora. Two types of experimental diets were formulated with 100% fish oil (FO) or 69% mixed oil (Mix). The results indicated that the Mix diet significantly improved the specific growth rate (SGR), weight gain (WG), feed efficiency ratio (FER), muscle tissue growth hormone receptor (GHR), insulin‐like growth factor 1 (IGF1) and insulin‐like growth factor 1 receptor (IGF1R) gene expression of the yellow drum (p < .05). No significant difference in the survival rate (SR), feed intake (FI), hepatosomatic index (HSI), viscerosomatic index (VSI), condition factor (CF), N3 PUFA percentage, liver antioxidant enzyme activities or pro‐inflammatory gene expression was observed between the two treatments. Therefore, terrestrial blend lipid from linseed and lard could be used as an alternative for dietary fish oil without compromising growth performance and immunity for the juvenile yellow drum.     

Effect of dietary vitamin C on growth performance,body composition and biochemical parameters of juvenile Chu's croaker (Nibea coibor)

Vitamin C is an essential micronutrient for normal physiological and immune functions of fish. However, its requirements and effects in Chu's croaker (Nibea coibor) are currently unknown. A 56‐day feeding trial was conducted to evaluate the optimal dietary vitamin C requirements based on its effects on growth performance, body composition and biochemical parameters in juvenile Chu's croaker (14.17 ± 0.1 g). Six isoproteic (450 g/kg crude protein) and isolipidic (100 g/kg crude lipid) diets were formulated to contain 2.24 (basal diet), 39.03, 85.01, 171.16, 356.49 and 715.46 mg/kg of vitamin C. The results showed that fish fed on 171.16 mg/kg vitamin C diet had the highest growth performance and feed utilization. Fish fed on the basal diet had higher malondialdehyde (MDA) content and lower activities of antioxidant enzymes in the serum and liver as compared with those fed on vitamin C diets. Polynomial analysis indicated that the optimal dietary vitamin C requirements of juvenile Chu's croaker were 102.28, 98.21, 150.26, 165.38, 71.46, 176.19, 84.84 and 103.78 mg/kg based on weight gain, specific growth rate, liver storage, muscle storage, liver MDA content, liver alanine aminotransferase activity, liver alkaline phosphatase activity and liver superoxide dismutase activity, respectively. We recommend an inclusion level in the range of 71.46–150.26 mg/kg vitamin C in the diets of juvenile Chu's croaker for optimum growth performance, liver function, antioxidant capacity and innate immunity functions.     

Effect of hydroxyproline supplementation on growth performance,body composition,amino acid profiles,blood‐biochemistry and collagen synthesis of juvenile chu's croaker (Nibea coibor)

Hydroxyproline (Hyp) may play an important role in collagen metabolism, as it is almost exclusively found in collagen. To examine this possibility, a 56‐day feeding trial was conducted to evaluate the effect of graded supplementaries of Hyp (0, 2.5, 5, 7.5, 10 and 12.5 g/kg dry diet weight) on growth performance, feed utilization, biochemical composition and collagen synthesis of chu's croaker, Nibea coibor (13.6 ± 0.28 g). The growth performance and feed utilization were improved with increased levels of dietary Hyp, reaching a maximum at 5 g/kg (diet H2) followed by a slight decrease. Polynomial regression analysis showed that the maximum specific growth rate (SGR) was attained at 7.177 g/kg dietary Hyp. Increasing levels of Hyp did not affect body composition of juvenile Nibea coibor. However, dietary Hyp supplementation significantly affected the levels of Leu, Ile, Met, Lys, Ser, Glu and Pro in muscle and the levels of Met, Lys, Glu, Pro and Gly in swim bladder. The serum calcium (SC) level decreased significantly with increasing levels of dietary Hyp. However, the triglycerides (TG) and total cholesterol (T‐CH) levels in serum and liver were relatively stable. Importantly, the acid‐soluble collagen (ASC) and total collagen (TC) levels in swim bladder increased significantly initially and then decreased after reaching a maximum. Polynomial regression analysis revealed that the maximum TC level in swim bladder was attained at 9.736 g/kg of dietary Hyp. Collectively, these results suggest that the appropriate provision of hydroxyproline is essential for maximal collagen synthesis and growth.     

Induction of sex reversal in blue drum (Nibea mitsukurii) and gynogenetic yellow drum (Nibea albiflora) by oral administration of letrozole

The objective of this study was to test the efficiency of an aromatase inhibitor, letrozole (LZ), to induce masculinization in blue drum (Nibea mitsukurii) and gynogenetic yellow drum (N. albiflora). Two experiments (Exp‐1 and Exp‐2) were performed to investigate the effect of LZ on the growth, sex ratio and gonad development in these two fishes. In Exp‐1, blue drum were treated with oral administration of letrozole at different doses (0, 1, 10, 100 mg/kg). In Exp‐2, gynogenetic yellow drum were orally administered different doses of LZ (0, 10, 100 mg/kg) and a dose of 10 mg/kg 17α‐methyltestosterone (MT). The treatments in both experiments were performed from 25 to 85 days post hatch (dph). As a result, all the LZ‐treated fish were phenotypic males despite of slower growth than the control during the treatment period. Free spermatozoa were observed from 150 dph onwards and the gonadosomatic index of the LZ‐treated fish did not differ significantly from that in the control at maturity. However, the sex‐reversed fish induced by MT exhibited a high proportion (33.4%) of testicular abnormalities. Our results showed that oral administration of LZ was efficient in inducing sex reversal in drums, and thereby facilitated their mono‐sex culture.     

Effect of Arthrospira supplementation on Oreochromis niloticus gut microbiota and flesh quality

Supplementing fish feed with Arthrospira platensis has several beneficial effects, such as improved growth, survival and feed conversion, but little is known about how it affects gut microbiota or flesh quality. In this trial, feed with 3% A. platensis was given to Oreochromis niloticus (SPR, spirulina supplemented) over a period of 50 days and their gut microbiota and flesh quality were compared with controls (CTRs, no supplementation). Average daily growth was higher in SPR fish (p < 0.05). Regarding microbiota, a total of 5,470,272 readings (individuals) were obtained from 16 gut samples (n = 8 CTR, n = 8 SPR), with a gamma diversity of 1,563 species but no significant differences were found between treatments. Among the high inter‐individual variability found, some individual microbiota were dominated by Proteobacteria (36.6%) and Fusobacteria (32.9%) and others more by Actinobacteria (37%). Regarding flesh quality, significant differences were observed in flesh colour post‐mortem (a*, b* and chroma) (p < 0.05), with higher reflectance in the range of 470–500 and 620–670 nm in SPR tilapia, which corresponds with the reflectance range of β‐carotene and phycocyanin. However, rigor mortis was resolved earlier in SPR fish (p < 0.05), which suggests a shorter shelf life. Adding 3% of A. platensis in O. niloticus diet can improve growth and flesh colour and does not appear to modify intestinal microbiota.     

Effects of natural dietary carotenoids on skin coloration of false Clownfish (Amphiprion ocellaris Cuvier, 1830)

This study was performed to evaluate the effect of dietary natural carotenoid sources on skin colour enhancement of false clownfish Amphiprion ocellaris. The juvenile fish (initial body wt. 0.30 g) were fed with four experimental diets including (a) commercial feed (reference diet), (b) moist feed, (c) sweet potato (potato diet) and (d) dried gut weed Enteromorpha sp. (gut weed diet). Sweet potato and gut weed were used to boost up β‐carotene levels in the diets. There was no significant difference in final weight (0.51 ± 0.02 – 0.61 ± 0.01 g) and length (2.80 ± 0.02 cm) of fishes among treatments (p > 0.05) after 8 weeks. The survival rate of the fish in all dietary treatment was greater than 89%. Principal component analysis results showed that fish fed potato and gut weed diets performed brighter colour in skin with more orangeness, body and accumulated β‐carotene levels were higher than those fish fed with reference diet. Indicator a* value for the redness of fish fed potato diet (16.18 ± 0.59) and gut weed (14.36 ± 2.14) was also higher than fish fed reference diet (10.92 ± 0.82). The result of this study provided key information for developing dietary colour enhancement of ornamental fish by using cost‐effective feed ingredients (potato and gut weed) as natural supplemental carotenoid sources.     

Effects of protein levels on growth,feed utilization,body composition,amino acid composition and physiology indices of juvenile chu's croaker,Nibea coibor

A 49‐day feeding trial was conducted to determine the optimal dietary protein level for juvenile chu's croaker Nibea coibor in sea cages. Five isocaloric diets were formulated to contain graded levels of protein: 360 (D36), 400 (D40), 440 (D44), 480 (D48) and 520 (D52) g kg^?1 diet. Each diet was randomly assigned to triplicate groups of 25 fish (mean 76.0 ± 0.8 g) in a total of 15 floating cages. Specific growth rate (SGR), protein retention efficiency (PRE), feed efficiency and feed intake were significantly higher at D44 and D48. Consumed protein (CP) obtained the maximum at D48, and protein efficiency ratio was significantly higher in fish fed with D44 and D48 compared with fish fed with D36 and D52. Hepatosomatic index and condition factor were not affected. Body protein and moisture content increased with increasing dietary protein, while lipid and ash content were not affected. Individual and total essential amino acids maximized at D44. Digestive enzymes and physiology indices in intestine, liver and plasma were differentially affected. Based on polynomial regression analysis of SGR, PRE and CP, we suggested that the optimum dietary protein level for juvenile chu's croaker was 444–478 g kg^?1 diet.     

Effects of dietary 60 g kg−1 dried distiller's grains in least‐cost practical diets on production and gut allochthonous bacterial composition of cage‐cultured fish: comparison among fish species with different natural food habits

Dried distiller's grain (DDG) is considered as an alternative ingredient of dietary feed due to its high contents of protein, fibre and fat. In this study, 60 g kg^‐1 of DDG was used to feed grass carp (Ctenopharyngodon idella), bluntnose black bream (Megalobrama amblycephala), gibel carp (Carassius gibelio) and black carp (Mylopharyngodon piceus) for 8 weeks, and its effect on fish production and gut allochthonous microbiota was investigated for the development of a suitable fish feed high in nutrients and low in cost for polyculture freshwater fish. DDG supplementation resulted in the less weight gain and higher feed conversion ratio of black carp (P < 0.05), but had no significant effects on other fish or parameters. PCR–denaturing gradient gel electrophoresis (DGGE) analysis indicated that all four fish species had some common and unique bacteria in their digestive tracts, and the gut microbiota of bluntnose black and gibel carp fed the control diet and DDG diets were very similar (Cs > 91%); of them, the total counts of intestinal bacteria studied by qPCR increased in grass carp (P < 0.05) and depressed in black carp (P < 0.05) when fed dietary DDG. Thus, we assumed that dietary DDG modulated production and gut microbiota of fish in a host‐specific way.     

Histological characterization of gonadal sex differentiation in Nibea albiflora

The aim of this study was to investigate the period of sex differentiation and the pattern of gonadal differentiation in Nibea albiflora using histological techniques. The fish were raised in temperature between 19.5 and 21.5°C. The promordial gonads formed at the posterior region of the abdomen and the fish remained undifferentiated till the 36 day post hatching (dph) with a total length (TL) of 21.20 ± 3.90 mm. Ovarian differentiation precedes testicular differentiation, as usual in teleost fish. Oocytes undergoing meiosis at zygotene (bouquet) and pachytene stages were firstly observed when the total length reached 33.05 ± 4.61 mm at 40 dph, and the formation of the ovarian cavity began simultaneously at 46 dph with total length of 49.40 ± 4.83 mm. When the mean total length reached 78.05 ± 5.24 mm (61 dph), ovarian cavity grew bigger, and oocytes were firstly detected. By contrast, signs of morphological differentiation in the presumptive testis are not observed until their total length reached 69.18 ± 5.26 mm at 55 dph. The unrestricted lobular organization of the testis is evident at the stage of TL = 78.05 ± 5.24 mm (61 dph). These findings indicate that N. albiflora is a differentiated gonochorist and the critical period of sex differentiation in N. albiflora occurs at the stage of TL = 21.20 ± 3.90 mm (36 dph).     

Weaning methods using formulated feeds for snakehead (Channa striata and Channa micropeltes) larvae

The culture of snakehead fish (Channa striata and Channa micropeltes) in Vietnam is limited, and snakehead culture has been banned in Cambodia, because traditional practices include capture of fingerlings from the wild as seed, as well as capture of small‐size (also known as trash fish or low‐value) fish. As hatchery breeding technology has improved, we investigated the optimal weaning practices for these two species. Both laboratory experiments and farm trials were conducted. For C. striata, the optimal weaning procedure is to begin at 17 days after hatch (dah) and wean the fish at 10% replacement of live feed with formulated feed per day. However, for C. micropeltes, the optimal procedure is to wait until 40 dah to begin weaning and then to wean the fish with a 10% replacement of live feed with formulated feed every 3 days. These results should enable farmers to domesticate snakehead culture in Vietnam and Cambodia and eliminate reliance on fish captured from the wild as both seed and feed.     

Effects of dietary proline on swim bladder collagen synthesis and its possible regulation by the TGF‐β/Smad pathway in spotted drum,Nibea diacanthus

Swim bladder is an ideal source of collagen production in fish for improved human health. Proline (Pro) is the main proteinogenic amino acid needed for collagen production. However, the effects of Pro supplementation on the swim bladder collagen synthesis have rarely been evaluated in fish. We determined the effects of dietary Pro supplementation on swim bladder collagen synthesis and its possible signalling pathway in spotted drum, Nibea diacanthus. A total of 450 N. diacanthus (100 ± 3.05 g) were randomly assigned into six treatments and fed with diets supplemented with different levels of Pro (0, 2.5, 5, 7.5, 10 and 12.5 g/kg of dry diet, hereafter P0, P1, P2, P3, P4 and P5, respectively) for 8 weeks. At the end, we evaluated collagen synthesis in swim bladder and the expression of genes related to TGF‐β/Smad pathway in the fish. Dietary Pro levels increased significantly the contents of crude protein, total collagen (TC) and the levels of some amino acids in swim bladder than the control diet (p < .05). The optimum amount of dietary Pro inclusion in diets for swim bladder collagen synthesis in N. diacanthus was 7.6 and 7.5 g/kg based on crude protein and TC in swim bladder, respectively. Dietary Pro levels increased significantly the proline 4‐hydroxylase (P4H) content in fish serum, swim bladder, muscle and liver tissues than control (p < .05). The relative expression of collagen type I alpha 1 (COL1A1), alpha 2 (COL1A2) and mothers against decapentaplegic homolog 2 (Smad2) genes in liver and swim bladder initially increased significantly as the concentration of Pro and later decreased (p < .05). Similarly, the relative expression of transforming growth factor beta (TGF‐β), P4Ha2 and P4Ha3 genes in the swim bladder increased significantly as dietary Pro levels increased (p < .05). Using K‐means clustering analysis, dietary proline partly promoted collagen accumulation in swim bladder through upregulation of Smad2 and TGF‐βRT genes. Taken together, Pro affected the collagen metabolism in swim bladder, probably by regulating the TGF‐β/Smad pathway, most likely via transient overexpression of Smad2 gene.     

Effects of conjugated linoleic acid on growth,body composition,antioxidant status,lipid metabolism and immunity parameters of juvenile Chu's croaker,Nibea coibor

A‐56 days feeding trial was conducted to evaluate the growth performance, feed utilization, biochemical composition, antioxidant status, lipid metabolism and immunity parameters of Chu's croaker Nibea coibor fed diets supplemented with different levels of conjugated linoleic acid (CLA): 0% (control), 0.5%, 1%, 1.5% and 2%. Each diet was randomly assigned to triplicate groups of 25 fish (initial body weight: 12.8 ± 0.1 g) in 15 floating cage. Weight gain (WG) and specific growth rate (SGR) were not significantly affected. 2% of dietary CLA led to significant lower lipid content in both whole body and liver (p < .05). Muscle lipid content was significantly lower in fish fed 1.5% and 2% CLA (p < .05). Saturated fatty acid (SFA) increased while monounsaturated fatty acid (MUFA) decreased with increasing dietary CLA in tissues (p < .05). Increasing CLA significantly promoted hepatic lipase (HL), lipoprotein lipase (LPL), serum lysozyme (LSZ) and immunoglobulin M (IgM) while decreased serum triglyceride (TG) in both liver and serum (p < .05). Liver Superoxide dismutase (SOD), catalase (CAT) and total antioxidant capacity (t‐AOC) increased while malondialdehyde (MDA) decreased (p < .05). Our study shown that Chu's croaker can successfully incorporate CLA in tissues up to 2% dietary lipid without growth inhibition, and that CLA increased fish quality due to the increased parameters of lipid metabolism, specific immunity, hepatic antioxidant and CLA accumulation, and the reduction of tissue fat deposition.     

Growth,feed utilization,body composition and swimming performance of giant croaker,Nibea japonica Temminck and Schlegel,fed at different dietary protein and lipid levels

A 50‐day feeding trial was conducted to examine the effects of dietary protein and lipid levels on growth, feed utilization, body composition and swimming performance of giant croaker, Nibea japonica. Fish (initial body weight 44.6 g ind⁻¹) were fed ten test diets which were formulated at 5 crude protein levels (360, 400, 440, 480 and 520 g kg⁻¹) and 2 crude lipid levels (90 and 150 g kg⁻¹). In addition, a raw fish diet (fillet of small yellow croaker) served as the reference. The weight gain (WG) increased, whereas the feed intake (FI) and feed conversion ratio (FCR) decreased, with increasing dietary protein level from 360 to 520 g kg⁻¹. At the same dietary protein level, no significant difference was found in the WG between fish fed the diets containing 90 or 150 g kg⁻¹ crude lipid. Fish fed the diet containing 480 g kg⁻¹ crude protein and 90 g kg⁻¹ crude lipid exhibited higher WG, nitrogen retention efficiency (NRE) and energy retention efficiency (ERE) but lower nitrogen wastes output (TNW). At the end of the feeding trial, the hepatosomatic index (HSI) and viscerosomatic index (VSI) decreased, whereas the body protein content increased, with increase in dietary protein level. The body lipid content was higher in fish fed at the 150 g kg⁻¹ lipid level than in fish fed at the 90 g kg⁻¹ lipid level. No significant difference was found in the maximum sustained swimming speed (MSS) between fish fed at different dietary protein and lipid levels. The WG, NRE, ERE and condition factor (CF) were higher, whereas the FI, FCR, HSI, VSI and TNW were lower, in fish fed the raw fish diet than in fish fed the diet containing 480 g kg⁻¹ crude protein and 90 g kg⁻¹ crude lipid. No significant difference was detected in the MSS between fish fed the raw fish diet and diet containing 480 g kg⁻¹ crude protein and 90 g kg⁻¹ crude lipid. The results of this study suggest that the suitable dietary crude protein and crude lipid levels are 480 g kg⁻¹ and 90 g kg⁻¹ for giant croaker reared in net pens.     

The effects of dietary vitamin C on growth performance,serum enzymes activities and resistance to Vibrio alginolyticus challenge of yellow drum Nibea albiflora

A 9‐week feeding trial was conducted to determine the optimal dietary vitamin C requirement and its effects on serum enzymes activities and bacterial resistance in the juvenile yellow drum Nibea albiflora (initial weight 33.2 ± 0.10 g). Six practical diets were formulated containing vitamin C 2.1, 45.3, 89.6, 132.4, 178.6 and 547.1 mg kg^?1 diet supplied as l ‐ascorbyl‐2‐monophosphate. The fish fed 547.1 mg kg^?1 diet showed a significantly higher survival than that fed 2.1 mg kg^?1 diet. The weight gains and specific growth rate of the fish fed 2.1 mg kg^?1 diet were significantly lower than those of the fish fed 89.6–547.1 mg kg^?1 diets. The liver vitamin C concentration firstly increased with increasing dietary vitamin C supply from 2.1 to 178.6 mg kg^?1 diet and then stabilized. The serum superoxide dismutase activities of the fish fed 547.1 mg kg^?1 diet were significantly lower than those of the fish fed 2.1–89.6 mg kg^?1 diet. The fish fed 2.1 mg kg^?1 diet had a significantly higher alkaline phosphatase activity than those in the other groups except the 45.3 mg kg^?1 group. Fish that received diets containing vitamin C at 547.1 mg kg^?1 had significantly higher nitro blue tetrazolium and lysozyme activity, and fish that received diets containing vitamin C at 45.3–547.1 mg kg^?1 exhibited resistance against Vibrio alginolyticus infection. The dietary vitamin C requirement of the juvenile yellow drum was established based on broken‐line model of weight gain to be 142.2 mg l ‐ascorbyl‐2‐monophosphate kg^?1 diet.     

Effects of kelp residues fermented with probiotics on the culture of sea cucumber,Apostichopus japonicus

The kelp aquaculture production in China is the largest in the world, and a large amount of kelp residue is produced by kelp processing. Kelp residues contain substantial quantities of crude fibre, protein, and residual alginic acid, and may be used as feedstuff for aquaculture animals. In this study, we used probiotics to ferment kelp residues to improve kelp nutrient content and then fed the fermented kelp to the sea cucumber, Apostichopus japonicus. To study the effect of fermented feed on sea cucumber, its growth performance, digestive enzyme activity, diversity of intestinal microbiota and water quality of the sea cucumber culture water were determined. Growth performance of sea cucumber fed with fermented feed significantly (p < .01) increased when compared with sea cucumber fed with formulated feed. Amylase, cellulose and alginase activities were significantly (p < .01) higher in the fermented feed group when compared with the formulated feed group. The total number and diversity of intestinal microbiota showed a significant increase in sea cucumbers fed with the fermented feed. The water quality of the fermented feed group showed much lower ammonia and nitrite (<0.050 mg/L) levels when compared with the formulated feed group. These results suggest that kelp residues fermented with probiotics enhance the growth, digestive enzyme activities and intestinal microbiota of sea cucumbers and improve the culture water quality. Fermented kelp residues are a new supplementary nutrient source for sea cucumbers and may be applicable to other animal aquacultures.     

Effect of Lactobacillus acidophilussupplementation on growth performances,digestive enzyme activities and gut histomorphology of striped catfish (Pangasianodon hypophthalmusSauvage, 1878) juveniles

The influence of dietary supplementation of Lactobacillus acidophilus, on growth performance, digestive enzyme activities, gut histomorphology and gut microflora were evaluated in juveniles striped catfish (Pangasianodon hypophthalmus). Five experimental diets were formulated by supplementing 0, 10³, 10⁵, 10⁷ and 10⁹ CFU/g L. acidophilus in fishmeal and casein‐based semi‐purified diet. Triplicate groups of striped catfish (21.69 ± 0.18 g) were stocked in 15 fiberglass tanks with stocking density of 25 individuals per tank and fed twice daily at 2.5% of the fish body weight for 12 weeks. Weight gain, specific growth rate, feed conversion ratio and the protein efficiency ratio were significantly higher in fish fed with 10⁵ and 10⁷CFU/g L. acidophilussupplemented diets compared with the other treatment groups. Compared with the control and fish fed low (10³ CFU/g) L. acidophilus supplementation, those fed with 10⁵ and 10⁷ CFU/g had significantly higher (p < 0.05) apparent protein digestibility. Inclusion of L. acidophilus at 10⁷ CFU/g diet significantly increased amylase, protease and lipase activities. Microscopic analysis showed that the villi length in both the anterior and posterior gut and microvilli length in the posterior gut increased significantly in fish fed L. acidophilus supplementation at 10⁵ and 10⁷ CFU/g of diet. The fish fed L. acidophilus supplemented diets significantly increased the total lactic acid bacteria counts in the gut of striped catfish compared with the control‐fed group. Based on gut histomorphology and growth performance, inclusion of L. acidophilus at 10⁵ CFU/g appears to have the most positive effect on striped catfish farming.     

Clostridium butyricum as probiotic for promoting growth performance,feed utilization,gut health and microbiota community of tilapia (Oreochromis niloticus × O. aureus)

The feeding trial was conducted to evaluate the potentials of Clostridium butyricum in the diet of tilapia. Fish (~14 g) were fed with basal diet supplemented with 0 (Control), 0.5 (C‐1), 1 (C‐2), 2 (C‐3), 4 (C‐4) and 8 (C‐5) g/kg commercial probiotic‐containing C. butyricum (1.5 × 10⁸ CFU/g) for 8 weeks. The results showed that weight gain significantly increased, and feed conversion ratio decreased in the C‐2, C‐3 and C‐4 groups (p < .05). The protein retention (except C‐1 group), lipid retention and apparent digestibility coefficient (ADC) of dry matter in probiotic supplementation groups were significantly enhanced, and ADC of protein in the C‐4 group was also improved (p < .05). The supplementation of probiotic significantly increased villus height in anterior intestines and reduced the numbers of intestinal Escherichia coli (p < .05). High‐throughput sequencing showed that top three phyla namely Planctomycetes in all probiotic‐containing groups, Proteobacteria in the C‐1 and C‐2 groups and Chloroflexi in the C‐3 group had higher level than the NC group. The cumulative mortality was reduced by dietary probiotic after challenging with Aeromonas hydrophila (p < .05). In conclusion, C. butyricum can be supplemented at 1–2 g/kg feed for promoting the growth, feed utilization, gut health and microbiota of tilapia.     

The effect of fresh seaweed and a formulated diet supplemented with seaweed on the growth and gonad quality of the collector sea urchin,Tripneustes gratilla,under farm conditions

This gonad enhancement study investigates the effect of different fresh and formulated feeds and feeding regimes on the growth and gonad quality of wild‐collected adult sea urchin, Tripneustes gratilla, under farm conditions for over 18 weeks. In the first 12 weeks (phase 1), urchins were fed fresh Ulva rigida (U); a 50:50 mixture of fresh U. rigida and Gracilaria gracilis (UG); fresh G. gracilis (G) and a formulated diet 20U (containing 20% U. rigida), and in the final 6 weeks (phase 2) of the study, diet was changed to a formulated feed (20U diet). By the end of phase 1, urchins fed the 20U diet produced gonads (50.72 ± 5.4 g) that were significantly heavier (p < .001) than the gonads of urchins fed the fresh seaweed diets (U, UG & G). By the end of phase 2, gonad weight of urchins in treatment groups UG‐20U and G‐20U were similar to those fed the 20U‐20U diet. Gonad colour of urchins in the G‐20U treatment became significantly lighter (ANOVA, p = .029) and poorer quality, compared with urchins in the U‐20U group. This gonad enhancement study, conducted on wild‐collected adult T. gratilla, has shown that a formulated feed (20U diet) can enhance gonad growth and produce commercially acceptable gonads. This farm‐based study supports previous findings from aquarium‐based studies by our group and indicates that short‐term sea urchin gonad enhancement can be carried out under farm conditions in South Africa.     

Prebiotics effect on growth performance,hepatic intermediary metabolism,gut microbiota and digestive enzymes of white sea bream (Diplodus sargus)

This study aimed at evaluating the effects of short‐chain fructooligosaccharides (scFOS), xylooligosaccharides (XOS) and galactooligosaccharides (GOS) on growth performance, hepatic metabolism, gut microbiota and digestive enzymes activities of white sea bream juveniles. Four diets were formulated: a control diet with fish meal (FM) and plant feedstuffs (PF) (30FM:70PF) and three test diets similar to control but supplemented with 10 g of scFOS, XOS or GOS per kilo diet, which were fed to fish during 12 weeks. Prebiotics had no effect on growth, feed efficiency or gut microbiota. Plasmatic triglycerides were lower in fish fed XOS than FOS and GOS diets. Malic enzyme activity was lower in fish fed XOS than FOS diet. Fish fed XOS diet had lower fatty acid synthetase (FAS), a key lipogenic enzyme and higher alanine aminotransferase activities. Fifteen days after the start of the trial, an enhancement of total alkaline protease, trypsin and lipase activities was observed in fish fed prebiotics, but such effect disappeared at 12 weeks. In conclusion, scFOS, XOS or GOS seem to have limited applicability in white sea bream feed.     

Microbial communities associated with early stages of intensively reared orange‐spotted grouper (Epinephelus coioides)

The gut microbiota plays key roles in the health and general welfare of fish larvae, the present study characterized the bacterial communities associated with grouper Epinephelus coioides larvae during a period of 22 days post hatch (DPH) in an intensive hatchery using both cultivation‐based and cultivation‐independent approaches. Both approaches confirmed that bacteria were present in the gut of larvae before and after the onset of exogenous feeding, and the number of cultiviable bacteria increased gradually from 2 DPH to 22 DPH. A more complex bacterial profile was present in larvae fed fertilizer oyster eggs for 4 days (8 DPH), probably as a result of the onset of exogenous feeding. Interestingly, similar internal microbiota were observed in larvae fed fertilized oyster eggs for 4 days (8 DPH) and rotifers for 2 weeks (22 DPH), although different microbial communities were present in the two feeds. This might suggest that the gut environment of E. coioides larvae selects for a common microbiota, which is more closely related with the rearing water than the two feeds. Therefore, bacterial community of the rearing water may play a critical role in the establishment of gut microbiota of fish larvae and more attention should be paid to its practical modulation by using probiotics. In addition, some potentially beneficial bacteria, such as Lactococcus spp., were the major components of the microbiota associated with fertilized oyster eggs, while these bacteria were not detected in larvae samples.