Fish cell line provide a useful tool for studies in virology and molecular biology.To establish a novel continuous marine fish cell line(EFF)from caudal fin tissue of brown-marbled grouper Epinephelus fuscoguttatus,it...Fish cell line provide a useful tool for studies in virology and molecular biology.To establish a novel continuous marine fish cell line(EFF)from caudal fin tissue of brown-marbled grouper Epinephelus fuscoguttatus,its susceptibility to the crustacean covert mortality nodavirus(CMNV)was evaluated.The primary cell cultures were initiated first by incomplete digestion of fin tissue blocks with dispase and collagenase,and then the explant was cultured in L-15 medium supplemented with 20%fetal bovine serum,10%grouper muscle extract,and 20-ng/mL growth factors of basic fibroblast growth factor(bFGF)and epidermal growth factor(EGF).The EFF cells were continuously passaged beyond 50 times in fibroblast-like morphology,and they grew well in L-15 medium supplemented with a lower concentration fetal bovine serum(10%)at 28℃ after passage 10,without muscle extract and the growth factors.In addition,their grouper origin was confirmed by chromosome analysis and cytochrome oxidase 1(CO 1)gene analysis of these EFF cells.Transfection experiment via lipofectamine 8000 indicated that the EFF cells had a high transfection potential with a transfection efficiency up to 32%.Cross-species viral susceptibility analysis showed that CMNV not only successfully infected the EFF cells as evidenced by obvious cytopathic effects like vacuolation,detachment and death of cells,but also multiplied in the EFF cells as indicated by the results of semi-quantitative RT-PCR in transmission electron microscopy.Therefore,the establishment of immortal EFF cell line provided a useful cell model for future works on the isolation,multiplication,and pathogenic mechanism of cross-species infection of CMNV as well as genetic manipulation.展开更多
This study aimed to study the development characteristics of Epinephelus fuscoguttatus at the embryo, larval, juvenile and young fish stages, thereby improving the breeding efficiency of E. fuscoguttatus. [Method] The...This study aimed to study the development characteristics of Epinephelus fuscoguttatus at the embryo, larval, juvenile and young fish stages, thereby improving the breeding efficiency of E. fuscoguttatus. [Method] The morphological characteristics and embryonic and post embryonic development of E. fuscoguttatus, including the occurrence and development of nervous system, digestive system and circulatory system, were observed and analyzed using eggs self-fertilized with artificially induced spawning in the breeding base of Hainan Province. [Result] The fertilized egg of E. fuscoguttatus is spherical, colorless transparent with single oil ball, and its diameter ranges from 0.83 to 0.94 mm. Based on the development process, the embryonic development of E. fuscoguttatus was divided into six stages, namely fertilized egg stage, cleavage stage, blastula stage, gastrula stage, idiosome formation stage and hatching stage. The entire embryonic development of E. fuscoguttatus required 20 h 47 min in the sea water with temperature of 28.4-31.9℃ salinity of 31-32 and pH of 8.4, and the total cumulative temperature was 455.361 ℃·h. The post embryonic development of E. fuscoguttatus was divided into larval stage, juvenile stage and young fish stage based on the features of the yolk-sac, second dorsal fin spine, pelvic fin spine, and body color. Accordingly, the larval stage was further classified into pre-larva and post-larva, with or without yolk-sac, respectively. The extension and shrinkage of the first pelvic fin spine and the second dorsal fin spine as well as the spinules on aforementioned spines were the most significant changes during the post embryonic development of E. fuscoguttatus. [Conclusion] This study will provide some basic data for industrial breeding of E. fuscoguttatus.展开更多
The impact of water temperature(24, 27, 30 and 33℃) and salinity(15, 20, 25, 30 and 33) on oxygen consumption(OCR) and ammonium excretion rate(AER) of ♀ Epinephelus fuscoguttatus ×♂ E. lanceolatus hybrid group...The impact of water temperature(24, 27, 30 and 33℃) and salinity(15, 20, 25, 30 and 33) on oxygen consumption(OCR) and ammonium excretion rate(AER) of ♀ Epinephelus fuscoguttatus ×♂ E. lanceolatus hybrid grouper juveniles(9.39 ± 0.07 g) were investigated under the fed and un-fed conditions. The results showed that the OCR and AER were significantly(P < 0.05) affected by temperature and salinity under both fed and un-fed conditions. When temperature was 24–33℃, the OCR and AER of fed hybrid grouper juveniles were 85.68%–129.52% and 125.78%–287.63%, respectively, higher than those of un-fed hybrid grouper juveniles. The O/N ratio, protein use(P_u), Q_(10)(respiration) and Q_(10)(excretion) of fed hybrid grouper juveniles were 14.43–24.01, 28.35% – 48.48%, 1.69 and 3.01, respectively. The O/N ratio, P_u, Q_(10)(respiration), Q_(10)(excretion) of un-fed hybrid grouper juveniles were 20.39 – 31.79, 22.16% – 34.34%, 1.23 and 1.17, respectively. When salinity was 15–33, the OCR and AER of fed hybrid grouper juveniles increased by 87.42% – 116.85% and 215.38% – 353.57%, respectively, over those of un-fed hybrid grouper juveniles. The O/N ratio and P_u of fed hybrid grouper juveniles were 14.48 – 17.78, 39.36% – 49.43%, respectively. The O/N ratio and Pu of un-fed hybrid grouper juveniles were 20.39 – 31.79 and 22.16% – 34.34%, respectively. The specific dynamic action(SDA) of hybrid grouper juveniles was mainly related to protein metabolism. The results had a guiding significance to the large-scale intensive aquaculture of hybrid grouper juveniles.展开更多
The effects of temperature and pH on embryonic development and survival rate of newly hatched larvae of Epinephelus fuscoguttatus were discussed by single factor experiment method. Under a salinity of 31‰, fertilized...The effects of temperature and pH on embryonic development and survival rate of newly hatched larvae of Epinephelus fuscoguttatus were discussed by single factor experiment method. Under a salinity of 31‰, fertilized eggs were incubated at different temperatures （17, 20, 23, 26, 29 （eontrol）, 32, 35 ℃ ） and different pH values （5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.4 （control）, 8.5, 9.0, 9.5 and 10.0）, the development rate of fertilized eggs, hatching rate, deformity rate, survival rate of newly hatched larvae, and optimum temperature and pH ranges for embryonic development were recorded and analyzed. The results showed that the incubation period y required by fertilized eggs was in remarkably negative correlation with temperature x, y =4 017. 3x-15277 , R2 =0. 971 3. At 29 ℃, the hatching rate was the highest, up to 92.00% ; this temperature was corresponding to the lowest deformity rate of newly hatched larvae （ 1.09% ） ; corresponding effective accumulative temperature was also the lowest （437. 410 ℃ . h）, and significantly different from other experimental groups （P 〈 0.05 ） ; but on the contrary, corresponding survival rates at 24 and 48 h reached, respectively, 88.03% and 82. 97% which were significantly different from other experimental groups （ P 〈 0.05）. At the pH of 8.4 in the CK group and the pH of 8.5 of one experimental group, the embryonic development rates were the highest; con-esponding hatching rates of fertilized eggs were the highest, respectively, of 88.33% and 83.33% which were significantly different from other groups （P 〈 0.05） ; and corresponding survival rates at 24 and 48 h were the highest, and reached 90.84% and 85.03%, respectively. In the pH range of 8.0 - 8.5, the deformity rate of newly hatched larvae was 0. It was indicating that for embryonic development of E. fuscoguttatus, the suitable water temperature value was in the range of 17 - 29 ℃, the optimum value was in the range of 23 -29℃; and the suitable pH value was in the range of 6.5 -9.5, and the optimum value was in the range of 8.0- 8.5.展开更多
The effects of fish meal (FM) replacement with graded level of soybean meal (SBM) on growth performance, feed utilization, survival rate and body composition were investigated in juvenile tiger grouper, Epinephelus fu...The effects of fish meal (FM) replacement with graded level of soybean meal (SBM) on growth performance, feed utilization, survival rate and body composition were investigated in juvenile tiger grouper, Epinephelus fuscoguttatus (initial body weight 13.9 ±0.65 g). Six experimental diets were formulated to contain 50% crude protein, 16% crude lipid and 365.8 kcal/100g feed with SBM replacing FM protein at 0% (SM 0), 10% (SM 0), 20% (SM 20), 30% (SM 30), 40% (SM 40) and 20% with phytase (SM 20P) replacement levels. At the end of the ten-week feeding trial, there were no significant differences detected in terms of growth performances (weight gain and specific growth rate), feed conversion ratio (FCR) and survival rates of fish fed with the control diet (SM 0), SM 20, SM 30 and SM 20P. Net protein utilization of fish fed SM 20P was higher than those fed with other diets suggesting an improved utilization of nutrients with phytase addition in the diet. Replacements of FM protein with SBM at 10% and 40% have resulted in significantly lower growth and poorer FCR than other replacement levels. Survival rates remain high (≥90%) throughout the trial. Whole-body proximate composition of the fish was significantly affected by the inclusion of SBM in the diets. It can be concluded that 20%-30% of FM protein replacement with SBM is recommended and addition of phytase in the SBM-based diet should be considered to improve nutrient utilization of tiger grouper juvenile.展开更多
The present study was conducted as a continuation of our previous study on the effects of fish meal replacement with soy bean meal, SBM in the diets of tiger grouper, Epinephelus fuscoguttatus which revealed the possi...The present study was conducted as a continuation of our previous study on the effects of fish meal replacement with soy bean meal, SBM in the diets of tiger grouper, Epinephelus fuscoguttatus which revealed the possible level of fish meal protein replacement of 30% with positive effect of phytase supplementation in one of the test diet. In the present study, all experimental diets with SBM inclusion (10%, 20%, 30%, 40% replacement levels) were supplemented with phytase and fed to juvenile tiger grouper with initial body weight of 44.57 ± 0.28 g for eight weeks. Apart from the effects on growth performance, diet utilization and fish whole-body composition, apparent digestibility coefficient of diets was also measured. Similar findings to our previous experiment were observed in the present study. Diet SM40P (40% replacement level) had produced the poorest growth performance and feed conversion ratio (FCR). However, it was observed that FCR of SM40P was improved when compared to SM40 (without phytase) in our previous experiment. Other than that, SBM-based diets were performed as good as the control diet in terms of growth, feed utilization and survival rate. Replacement of fish meal did not significantly influence body composition, except for whole body lipid content in SM40P. In general, dry matter and protein apparent digestibility coefficients (ADC) of the test diets were influenced by SBM inclusion in the diets. However, crude lipid ADC was very high in all dietary treatments indicating efficient utilization of dietary lipid by juvenile tiger grouper. The findings of the present study confirmed that SBM can be used to replace FM at a level of up to 30% and supplementation of phytase in SBM-based diets is recommended to improve the diet performance.展开更多
Several reports have revealed the vital role that probiotics play in fish growth and health.However,few works are available for host gut-derived probiotics on the growth,immunity,and gut microbiota of fish,especially ...Several reports have revealed the vital role that probiotics play in fish growth and health.However,few works are available for host gut-derived probiotics on the growth,immunity,and gut microbiota of fish,especially in hybrid grouper (♀Epinephelus fuscoguttatus×♂Epinephelus lanceolatus) due to their isolation difficulty and functional verification.This study aimed at assessing 3 host gut-derived Bacillus species?effects on the growth,immune and antioxidant-biochemical responses,haematological parameters,intestinal morphology,immune-related gene expression,gut microbiota,and disease resistance against Vibrio harveyi in hybrid grouper.A total of 480 hybrid grouper (initial weight=9.03±0.02 g) were randomly allotted into 4 groups,namely,the group fed a basal diet without probiotic inclusion (control,B0),the group fed the basal diet with Bacillus velezensis GPSAK4 (BV),the group fed the basal diet with Bacillus subtilis GPSAK9 (BS),and the group fed the basal diet with Bacillus tequilensis GPSAK2 (BT) strains at 1.0×10^(9)CFU/g.After a 6-week feeding trial,the results revealed significant improvements (P<0.05) in the growth performance,whole fish-body proximate composition,blood haematological parameters,serum,liver,and intestinal biochemical indexes,intestinal morphology,and protection against V.harveyi pathogen in the probiotic-treated groups compared with the untreated.Additionally,the expressions of intestinal tight junction genes (occludin and ZO1),pro-and anti-inflammatory genes,including IL1β,IL6,IL8,TNFa,MyD88,IL10,and TGFβ,were upregulated (P<0.05) after Bacillus species administration.Host gut-derived Bacillus supplementation shaped the gut microbiota by significantly increasing (P<0.05) the relative abundance of Proteobacteria,Bacteroidetes,Actinobacteria (except the BS group),Acidobacteria(except the BT group),Cyanobacteria (except the BV and BT groups),and Verrucomicrobia phyla,as well as known beneficial genera (Romboutsia,Turicibacter,Epulopiscium,Clostridium_sensu_stricto 1 and 13,Lactobacillus,and Bacillus),but significantly decreased (P<0.05) the abundance of Firmicutes,Chloroflexi,and Fusobacteria phyla,and purported pathogenic genera (Staphylococcus and Photobacterium) compared with the control group.Collectively,the results suggest that B.velezensis GPSAK4,B.subtilis GPSAK9(especially this strain),B.tequilensis GPSAK2 dietary supplementation at 1.0×10^(9)CFU/g has positive effects on the intestinal health of hybrid grouper via microbial composition modulation,thus enhancing the assimilation and absorption of nutrients to boost fish growth,immunity,and disease resistance.展开更多
Due to diminishing fish meal(FM)supplies,superb protein(PRO)sources are needed for aquaculture,such as soy-based PRO.However,these can cause enteritis and even intestinal injury in fish when used at high proportions i...Due to diminishing fish meal(FM)supplies,superb protein(PRO)sources are needed for aquaculture,such as soy-based PRO.However,these can cause enteritis and even intestinal injury in fish when used at high proportions in feed.This research examines the effects of substituting soybean protein concentrate(SPC)for FM on the growth performance and intestinal balance of pearl gentian groupers and investigates the mechanism of SPC-induced enteritis.Experimental fish(n=720)were fed 1 of 3 following diets:(1)a 50%FM diet(control),(2)a diet with 20%of the FM substituted with SPC(group SPC20),and(3)a 40%SPC-substituted diet(SPC40).Fish were fed for 10 wk iso-nitrogenous(50%PRO)and iso-lipidic(10%lipid)diets.Groups SPC20 and SPC40 showed significantly lower developmental performance and intestinal structures than control.Group SPC40 had significantly higher expressions of pro-inflammatory-related genes,such as interleukin 1β(IL1β),IL12,IL17 and tumor necrosis factorαand significantly lower expressions of anti-inflammatory-related genes,such as IL5,IL10 and transforming growth factorβ1.Biochemical and 16S high-throughput sequencing showed that the abundance and functions of intestinal flora in group SPC40 were significantly affected(P<0.05),and there were significant correlations between operational taxonomic unit abundance variations and inflammatory gene expressions at genus level(P<0.05).The second-and third-generation full-length transcriptome sequence was used to analyze the mechanism of SPC-induced enteritis in pearl gentian groupers,which showed that enteritis induced by SPC may be caused by disturbances to intestinal immune function induced by an imbalance in intestinal nutrition and metabolism,such as the intestinal immunity network for IgA production pathway.However,it remains unclear as to which intestinal immune or nutritional imbalance is most important in enteritis development.This study provides a basis for further research into soy PRO-related enteritis in fish.展开更多
Groupers are dispersed worldwide in the tropical and subtropical waters.They are prized in the live reef fish trade,making them candidates with high market value and consumer demand.In the Asian-Pacific region,the bro...Groupers are dispersed worldwide in the tropical and subtropical waters.They are prized in the live reef fish trade,making them candidates with high market value and consumer demand.In the Asian-Pacific region,the brown-marbled grouper(Epinephelus fuscoguttatus)is widely adapted as an aquaculture species.However,health management remains a major concern in the stressful intensive culture process.The present review discusses techniques and current knowledge in the cultivation of E.fuscoguttatus towards healthy fish growth focusing on aspects of immunity,feed,and husbandry.Understanding how the fish immune system responds during infections provides insights into the intricate ways fish resist pathogens.This information is helpful when developing vaccination strategies or immunostimulant compounds to strengthen fish immunity.Feeds that are formulated according to the needs of the fish ensure optimal growth and using suitable alternative ingredients may lower production cost without compromising fish health.Good husbandry practices contribute to a favourable environment for the fish to grow,while interspecific hybridization may be a convenient approach to generate hardier species.Nevertheless,it has been noted that research in E.fuscoguttatus is relatively scarce in contrast to the closely-related orange-spotted grouper(E.coioides),which is used in the present article as a reference.Ultimately,the identified gaps in knowledge between the two species warrant species-specific research in E.fuscoguttatus to promote fish health and ensure continued success in aquaculture.展开更多
基金Supported by the Key Research&Development Program of Shandong Province(No.2023 CXGC 010710)the National Natural Science Foundation of China(No.32273116)the Fundamental Research Funds for the Central Universities(No.202261023)。
文摘Fish cell line provide a useful tool for studies in virology and molecular biology.To establish a novel continuous marine fish cell line(EFF)from caudal fin tissue of brown-marbled grouper Epinephelus fuscoguttatus,its susceptibility to the crustacean covert mortality nodavirus(CMNV)was evaluated.The primary cell cultures were initiated first by incomplete digestion of fin tissue blocks with dispase and collagenase,and then the explant was cultured in L-15 medium supplemented with 20%fetal bovine serum,10%grouper muscle extract,and 20-ng/mL growth factors of basic fibroblast growth factor(bFGF)and epidermal growth factor(EGF).The EFF cells were continuously passaged beyond 50 times in fibroblast-like morphology,and they grew well in L-15 medium supplemented with a lower concentration fetal bovine serum(10%)at 28℃ after passage 10,without muscle extract and the growth factors.In addition,their grouper origin was confirmed by chromosome analysis and cytochrome oxidase 1(CO 1)gene analysis of these EFF cells.Transfection experiment via lipofectamine 8000 indicated that the EFF cells had a high transfection potential with a transfection efficiency up to 32%.Cross-species viral susceptibility analysis showed that CMNV not only successfully infected the EFF cells as evidenced by obvious cytopathic effects like vacuolation,detachment and death of cells,but also multiplied in the EFF cells as indicated by the results of semi-quantitative RT-PCR in transmission electron microscopy.Therefore,the establishment of immortal EFF cell line provided a useful cell model for future works on the isolation,multiplication,and pathogenic mechanism of cross-species infection of CMNV as well as genetic manipulation.
基金Supported by Special Project of Hainan Provincial Engineering Research Center(GCZX2013005)Technology Research and Development Project of Hainan Provincial Research Institutes(KYYS-2014-59,KYYS-2013-24,11-20410-0010)Applied Technology Research and Development and Demonstration Project of Hainan Province(ZDXM2015025)~~
文摘This study aimed to study the development characteristics of Epinephelus fuscoguttatus at the embryo, larval, juvenile and young fish stages, thereby improving the breeding efficiency of E. fuscoguttatus. [Method] The morphological characteristics and embryonic and post embryonic development of E. fuscoguttatus, including the occurrence and development of nervous system, digestive system and circulatory system, were observed and analyzed using eggs self-fertilized with artificially induced spawning in the breeding base of Hainan Province. [Result] The fertilized egg of E. fuscoguttatus is spherical, colorless transparent with single oil ball, and its diameter ranges from 0.83 to 0.94 mm. Based on the development process, the embryonic development of E. fuscoguttatus was divided into six stages, namely fertilized egg stage, cleavage stage, blastula stage, gastrula stage, idiosome formation stage and hatching stage. The entire embryonic development of E. fuscoguttatus required 20 h 47 min in the sea water with temperature of 28.4-31.9℃ salinity of 31-32 and pH of 8.4, and the total cumulative temperature was 455.361 ℃·h. The post embryonic development of E. fuscoguttatus was divided into larval stage, juvenile stage and young fish stage based on the features of the yolk-sac, second dorsal fin spine, pelvic fin spine, and body color. Accordingly, the larval stage was further classified into pre-larva and post-larva, with or without yolk-sac, respectively. The extension and shrinkage of the first pelvic fin spine and the second dorsal fin spine as well as the spinules on aforementioned spines were the most significant changes during the post embryonic development of E. fuscoguttatus. [Conclusion] This study will provide some basic data for industrial breeding of E. fuscoguttatus.
基金supported by the project of National Science and Technology Supporting Plan (No. 2011BAD13B04)
文摘The impact of water temperature(24, 27, 30 and 33℃) and salinity(15, 20, 25, 30 and 33) on oxygen consumption(OCR) and ammonium excretion rate(AER) of ♀ Epinephelus fuscoguttatus ×♂ E. lanceolatus hybrid grouper juveniles(9.39 ± 0.07 g) were investigated under the fed and un-fed conditions. The results showed that the OCR and AER were significantly(P < 0.05) affected by temperature and salinity under both fed and un-fed conditions. When temperature was 24–33℃, the OCR and AER of fed hybrid grouper juveniles were 85.68%–129.52% and 125.78%–287.63%, respectively, higher than those of un-fed hybrid grouper juveniles. The O/N ratio, protein use(P_u), Q_(10)(respiration) and Q_(10)(excretion) of fed hybrid grouper juveniles were 14.43–24.01, 28.35% – 48.48%, 1.69 and 3.01, respectively. The O/N ratio, P_u, Q_(10)(respiration), Q_(10)(excretion) of un-fed hybrid grouper juveniles were 20.39 – 31.79, 22.16% – 34.34%, 1.23 and 1.17, respectively. When salinity was 15–33, the OCR and AER of fed hybrid grouper juveniles increased by 87.42% – 116.85% and 215.38% – 353.57%, respectively, over those of un-fed hybrid grouper juveniles. The O/N ratio and P_u of fed hybrid grouper juveniles were 14.48 – 17.78, 39.36% – 49.43%, respectively. The O/N ratio and Pu of un-fed hybrid grouper juveniles were 20.39 – 31.79 and 22.16% – 34.34%, respectively. The specific dynamic action(SDA) of hybrid grouper juveniles was mainly related to protein metabolism. The results had a guiding significance to the large-scale intensive aquaculture of hybrid grouper juveniles.
基金Supported by Special Fund for Construction of Hainan Key Laboratory and Engineering Technical Research Center(gczx2013005)Special Fund for Technology Development by Scientific Research Institutes of Hainan Province(KYYS-2014-59)+1 种基金Special Fund for Application Technology Development and Demonstration(ZDXM2015025)Hainan Provincial-level Department Budget Project
文摘The effects of temperature and pH on embryonic development and survival rate of newly hatched larvae of Epinephelus fuscoguttatus were discussed by single factor experiment method. Under a salinity of 31‰, fertilized eggs were incubated at different temperatures （17, 20, 23, 26, 29 （eontrol）, 32, 35 ℃ ） and different pH values （5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.4 （control）, 8.5, 9.0, 9.5 and 10.0）, the development rate of fertilized eggs, hatching rate, deformity rate, survival rate of newly hatched larvae, and optimum temperature and pH ranges for embryonic development were recorded and analyzed. The results showed that the incubation period y required by fertilized eggs was in remarkably negative correlation with temperature x, y =4 017. 3x-15277 , R2 =0. 971 3. At 29 ℃, the hatching rate was the highest, up to 92.00% ; this temperature was corresponding to the lowest deformity rate of newly hatched larvae （ 1.09% ） ; corresponding effective accumulative temperature was also the lowest （437. 410 ℃ . h）, and significantly different from other experimental groups （P 〈 0.05 ） ; but on the contrary, corresponding survival rates at 24 and 48 h reached, respectively, 88.03% and 82. 97% which were significantly different from other experimental groups （ P 〈 0.05）. At the pH of 8.4 in the CK group and the pH of 8.5 of one experimental group, the embryonic development rates were the highest; con-esponding hatching rates of fertilized eggs were the highest, respectively, of 88.33% and 83.33% which were significantly different from other groups （P 〈 0.05） ; and corresponding survival rates at 24 and 48 h were the highest, and reached 90.84% and 85.03%, respectively. In the pH range of 8.0 - 8.5, the deformity rate of newly hatched larvae was 0. It was indicating that for embryonic development of E. fuscoguttatus, the suitable water temperature value was in the range of 17 - 29 ℃, the optimum value was in the range of 23 -29℃; and the suitable pH value was in the range of 6.5 -9.5, and the optimum value was in the range of 8.0- 8.5.
文摘The effects of fish meal (FM) replacement with graded level of soybean meal (SBM) on growth performance, feed utilization, survival rate and body composition were investigated in juvenile tiger grouper, Epinephelus fuscoguttatus (initial body weight 13.9 ±0.65 g). Six experimental diets were formulated to contain 50% crude protein, 16% crude lipid and 365.8 kcal/100g feed with SBM replacing FM protein at 0% (SM 0), 10% (SM 0), 20% (SM 20), 30% (SM 30), 40% (SM 40) and 20% with phytase (SM 20P) replacement levels. At the end of the ten-week feeding trial, there were no significant differences detected in terms of growth performances (weight gain and specific growth rate), feed conversion ratio (FCR) and survival rates of fish fed with the control diet (SM 0), SM 20, SM 30 and SM 20P. Net protein utilization of fish fed SM 20P was higher than those fed with other diets suggesting an improved utilization of nutrients with phytase addition in the diet. Replacements of FM protein with SBM at 10% and 40% have resulted in significantly lower growth and poorer FCR than other replacement levels. Survival rates remain high (≥90%) throughout the trial. Whole-body proximate composition of the fish was significantly affected by the inclusion of SBM in the diets. It can be concluded that 20%-30% of FM protein replacement with SBM is recommended and addition of phytase in the SBM-based diet should be considered to improve nutrient utilization of tiger grouper juvenile.
文摘The present study was conducted as a continuation of our previous study on the effects of fish meal replacement with soy bean meal, SBM in the diets of tiger grouper, Epinephelus fuscoguttatus which revealed the possible level of fish meal protein replacement of 30% with positive effect of phytase supplementation in one of the test diet. In the present study, all experimental diets with SBM inclusion (10%, 20%, 30%, 40% replacement levels) were supplemented with phytase and fed to juvenile tiger grouper with initial body weight of 44.57 ± 0.28 g for eight weeks. Apart from the effects on growth performance, diet utilization and fish whole-body composition, apparent digestibility coefficient of diets was also measured. Similar findings to our previous experiment were observed in the present study. Diet SM40P (40% replacement level) had produced the poorest growth performance and feed conversion ratio (FCR). However, it was observed that FCR of SM40P was improved when compared to SM40 (without phytase) in our previous experiment. Other than that, SBM-based diets were performed as good as the control diet in terms of growth, feed utilization and survival rate. Replacement of fish meal did not significantly influence body composition, except for whole body lipid content in SM40P. In general, dry matter and protein apparent digestibility coefficients (ADC) of the test diets were influenced by SBM inclusion in the diets. However, crude lipid ADC was very high in all dietary treatments indicating efficient utilization of dietary lipid by juvenile tiger grouper. The findings of the present study confirmed that SBM can be used to replace FM at a level of up to 30% and supplementation of phytase in SBM-based diets is recommended to improve the diet performance.
基金supported financially by the General Program of Natural Science Foundation of Guangdong Province(2021A1515011165)the Department of Education of Guangdong Province (2021ZDZX4005)+2 种基金the National Natural Science Foundation of China (31972808)the Research and Demonstration of Precision Functional Compound Feed Technology of Major Cultured Fishes and Shrimps in South China (2021B0202050002)the China Agriculture Research System of MOF and MARA (CARS-47)。
文摘Several reports have revealed the vital role that probiotics play in fish growth and health.However,few works are available for host gut-derived probiotics on the growth,immunity,and gut microbiota of fish,especially in hybrid grouper (♀Epinephelus fuscoguttatus×♂Epinephelus lanceolatus) due to their isolation difficulty and functional verification.This study aimed at assessing 3 host gut-derived Bacillus species?effects on the growth,immune and antioxidant-biochemical responses,haematological parameters,intestinal morphology,immune-related gene expression,gut microbiota,and disease resistance against Vibrio harveyi in hybrid grouper.A total of 480 hybrid grouper (initial weight=9.03±0.02 g) were randomly allotted into 4 groups,namely,the group fed a basal diet without probiotic inclusion (control,B0),the group fed the basal diet with Bacillus velezensis GPSAK4 (BV),the group fed the basal diet with Bacillus subtilis GPSAK9 (BS),and the group fed the basal diet with Bacillus tequilensis GPSAK2 (BT) strains at 1.0×10^(9)CFU/g.After a 6-week feeding trial,the results revealed significant improvements (P<0.05) in the growth performance,whole fish-body proximate composition,blood haematological parameters,serum,liver,and intestinal biochemical indexes,intestinal morphology,and protection against V.harveyi pathogen in the probiotic-treated groups compared with the untreated.Additionally,the expressions of intestinal tight junction genes (occludin and ZO1),pro-and anti-inflammatory genes,including IL1β,IL6,IL8,TNFa,MyD88,IL10,and TGFβ,were upregulated (P<0.05) after Bacillus species administration.Host gut-derived Bacillus supplementation shaped the gut microbiota by significantly increasing (P<0.05) the relative abundance of Proteobacteria,Bacteroidetes,Actinobacteria (except the BS group),Acidobacteria(except the BT group),Cyanobacteria (except the BV and BT groups),and Verrucomicrobia phyla,as well as known beneficial genera (Romboutsia,Turicibacter,Epulopiscium,Clostridium_sensu_stricto 1 and 13,Lactobacillus,and Bacillus),but significantly decreased (P<0.05) the abundance of Firmicutes,Chloroflexi,and Fusobacteria phyla,and purported pathogenic genera (Staphylococcus and Photobacterium) compared with the control group.Collectively,the results suggest that B.velezensis GPSAK4,B.subtilis GPSAK9(especially this strain),B.tequilensis GPSAK2 dietary supplementation at 1.0×10^(9)CFU/g has positive effects on the intestinal health of hybrid grouper via microbial composition modulation,thus enhancing the assimilation and absorption of nutrients to boost fish growth,immunity,and disease resistance.
基金This study received assistance from the National Key R&D Program of China(2019YFD0900200).
文摘Due to diminishing fish meal(FM)supplies,superb protein(PRO)sources are needed for aquaculture,such as soy-based PRO.However,these can cause enteritis and even intestinal injury in fish when used at high proportions in feed.This research examines the effects of substituting soybean protein concentrate(SPC)for FM on the growth performance and intestinal balance of pearl gentian groupers and investigates the mechanism of SPC-induced enteritis.Experimental fish(n=720)were fed 1 of 3 following diets:(1)a 50%FM diet(control),(2)a diet with 20%of the FM substituted with SPC(group SPC20),and(3)a 40%SPC-substituted diet(SPC40).Fish were fed for 10 wk iso-nitrogenous(50%PRO)and iso-lipidic(10%lipid)diets.Groups SPC20 and SPC40 showed significantly lower developmental performance and intestinal structures than control.Group SPC40 had significantly higher expressions of pro-inflammatory-related genes,such as interleukin 1β(IL1β),IL12,IL17 and tumor necrosis factorαand significantly lower expressions of anti-inflammatory-related genes,such as IL5,IL10 and transforming growth factorβ1.Biochemical and 16S high-throughput sequencing showed that the abundance and functions of intestinal flora in group SPC40 were significantly affected(P<0.05),and there were significant correlations between operational taxonomic unit abundance variations and inflammatory gene expressions at genus level(P<0.05).The second-and third-generation full-length transcriptome sequence was used to analyze the mechanism of SPC-induced enteritis in pearl gentian groupers,which showed that enteritis induced by SPC may be caused by disturbances to intestinal immune function induced by an imbalance in intestinal nutrition and metabolism,such as the intestinal immunity network for IgA production pathway.However,it remains unclear as to which intestinal immune or nutritional imbalance is most important in enteritis development.This study provides a basis for further research into soy PRO-related enteritis in fish.
基金Our research was supported by Malaysian Ministry of Education(MOE)Higher Institution Centre of Excellence(HICoE)Grant No.6369100Malaysian Ministry of Science,Technology and Innovation(MOSTI)EScience Fund No.5450576.
文摘Groupers are dispersed worldwide in the tropical and subtropical waters.They are prized in the live reef fish trade,making them candidates with high market value and consumer demand.In the Asian-Pacific region,the brown-marbled grouper(Epinephelus fuscoguttatus)is widely adapted as an aquaculture species.However,health management remains a major concern in the stressful intensive culture process.The present review discusses techniques and current knowledge in the cultivation of E.fuscoguttatus towards healthy fish growth focusing on aspects of immunity,feed,and husbandry.Understanding how the fish immune system responds during infections provides insights into the intricate ways fish resist pathogens.This information is helpful when developing vaccination strategies or immunostimulant compounds to strengthen fish immunity.Feeds that are formulated according to the needs of the fish ensure optimal growth and using suitable alternative ingredients may lower production cost without compromising fish health.Good husbandry practices contribute to a favourable environment for the fish to grow,while interspecific hybridization may be a convenient approach to generate hardier species.Nevertheless,it has been noted that research in E.fuscoguttatus is relatively scarce in contrast to the closely-related orange-spotted grouper(E.coioides),which is used in the present article as a reference.Ultimately,the identified gaps in knowledge between the two species warrant species-specific research in E.fuscoguttatus to promote fish health and ensure continued success in aquaculture.
