河北省海水养殖凡纳滨对虾(Litopenaeus vannamei)养殖尾水监测对比研究

以河北省黄骅地区海水工厂化和池塘养殖凡纳滨对虾尾水为研究对象,检测这两种养殖模式下不同养殖阶段所排放的养殖尾水中化学需氧量(COD)、无机氮(DIN)及活性磷酸盐(DIP)等指标的变化,利用单因子污染指数(P i)和内梅罗综合指数(P)对养殖尾水状况进行了评价。结果表明,尾水的主要污染因子为DIN与DIP;主要污染物浓度均值工厂化养殖模式高于池塘养殖模式,且工厂化养殖尾水主要污染物均超过《河北省海水养殖尾水污染物排放标准》一级排放限值;养殖尾水主要污染物浓度均随养殖时间的延长而增加;两种模式下养殖尾水的内梅罗综合指数P值均处于严重污染状等级。

A Transcriptome Analysis of Neural Tissue of Litopenaeus vannamei After Acute Exposure to Alexandrium pacificum

Alexandrium pacificum(A.pacificum)is a typical paralytic shellfish poisonous dinoflagellate.Harmful algal blooms(HABs)caused by this species can bring serious environmental problems and economic losses to the aquaculture industry.In this study,transcriptome sequencing and analyses were performed on the neural tissue of Litopenaeus vannamei(L.vannamei)after acute exposure to A.pacificum disrupted solution for 72 h,and differentially expressed genes(DEGs)were identified.The results showed that,compared with the control samples,300 DEGs were identified in the experimental group,of which 194 were up-regulated,and 106 down-regulated.The gene ontology(GO)functional enrichment analysis showed that DEGs were significantly enriched in the cortical cytoskeleton organization,troponin complex,amylo-alpha-1,6-glucosidase and thymidine phosphorylase.Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis found that DEGs were mainly enriched in the oxidative phosphorylation process,intercellular tight junctions and mitophagy.The results showed that the proteoglycans,signaling pathways,and various metabolic processes that regulate cell proliferation,differentiation,and apoptosis all played an essential role in the response of L.vannamei to A.pacificum toxins.

Insight into the quality deterioration induced by microorganisms in Litopenaeus vannamei during superchilling storage

Quality deterioration induced by microbial community in Litopenaeus vannamei during superchilling storage were demonstrated in this study.In this study,6 microorganisms were identified as the biomarkers by 16S rDNA sequencing and 71 differential metabolites were identified by untargeted metabolomics.The correlation analysis between microorganisms and differential metabolites revealed that Oceanicaulis,Aliihoeflea,Prauserella,Chelativorans and Pseudoalteromonas had the potential to be the important microorganisms of deterioration of superchilled shrimp.Moreover,the spearman correlation analysis revealed that lightness value had highly significant correlation with Shewanella(P<0.01),and thiobarbituric acid content,total viable count value,total sulfhydryl content,trichloroacetic acid-soluble peptide and Ca2+-ATPase activity showed significantly correlation with Oceanicaulis,Prauserella,Chelativorans,Aliihoefl ea and Pseudoalteromonas(P<0.05).This study offers a better understanding for mechanisms behind quality degradation induced by microorganisms,which may help the quality maintaining of Pacific white shrimp during superchilling storage.

串联养殖模式下的凡纳滨对虾(Litopenaeus vannamei)与缢蛏(Sinonovacula constricta)生长特性、消化免疫及水生态效应

虾蛏串养模式作为海水围塘养殖坚持可持续发展、绿色发展的模式,已在福建沿海地区推广,取得明显效果,但目前适宜于该模式下的虾蛏数量配比尚不清楚。在实验室内,通过设立LP组(虾蛏数量配比1︰3)、MP组(1︰5)、HP组(1︰7),模拟虾蛏串联养殖模式,对虾和缢蛏分别投喂颗粒饲料、微绿球藻和角毛藻,并对虾蛏生长特性、养殖环境水质因子、消化与免疫指标、环境与蛏内脏团微生物群落进行探究。结果表明,LP组和MP组虾蛏增重率和特定生长率显著高于HP组。LP组和MP组对虾增重率分别为43.88%、41.91%,缢蛏增重率分别为29.30%、29.40%。水质营养盐浓度开始总体升高达到峰值后总体下降。虾蛏对环境反应不同,缢蛏中淀粉酶和过氧化氢酶活性变化显著,对虾则是胰蛋白酶和超氧化物歧化酶活性变化显著。RDA分析表明,总氮、活性磷酸盐是影响微生物群落关键因素。16S rRNA分析表明,虾蛏不同数量配比对养殖环境以及缢蛏内脏团有较大影响,与LP组相比,MP组和HP组微生物种类更多,且参与氮磷循环的微生物丰度增加。当虾蛏数量配比为1︰3或者1︰5时,虾蛏能良好生长且串养系统养殖环境能起到改善作用。研究结果有助于优化虾蛏串养模式,对今后虾蛏串养模式健康发展具有重要意义。

Transcriptomes of Litopenaeus vannamei reveal modulation of antioxidant system induced by dietary archaeal carotenoids

Oxidative stress induced by factors such as ammonia nitrogen has become a major issue in shrimp farming.The effects of carotenoids on the growth and antioxidant capability of Litopenaeus vannamei juveniles were investigated in this study using dietary archaeal carotenoids supplementation.For four weeks,shrimp were given diets containing 0 mg/kg(Ctrl)and 55.98 mg/kg(Car)archaeal carotenoids.Dietary archaeal carotenoids significantly enhanced the astaxanthin content in shrimp muscles and carapaces,as well as the superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)activity(P<0.05).The malonaldehyde(MDA)content in Car group significantly decreased(P<0.05).The transcriptome analysis was conducted to determine the molecular processes in response to archaeal carotenoids supplementation.A total of 1536 differentially expressed genes(DEGs)were detected,including 538 upregulated DEGs and 998 downregulated DEGs.GO functional enrichment analysis between Ctrl and Car indicated that 26 GO terms including extracellular region,metabolic process,and proteolysis were enriched.The KEGG pathway enrichment analysis revealed that the amino sugar and nucleotide sugar metabolism,cysteine and methionine metabolism,glycine serine and threonine metabolism,and amino acid biosynthesis were enriched.Archaeal carotenoids influenced the expression of several important genes involved in reactive oxygen species(ROS)generation,Nrf2 signaling,and antioxidant enzymes.Seven DEGs were chosen to confirm the RNA-Seq data using qRT-PCR.The genes and pathways discovered in this work assist to elucidate the molecular processes through which archaeal carotenoid enhances L.vannamei antioxidative system.

The dissolution of total suspended solids and treatment strategy of tailwater in a Litopenaeus vannamei recirculating aquaculture system

In recirculating aquaculture systems(RASs),the effective treatment of aquaculture tailwater is essential to maintain the health of the RAS.This study investigated the optimal time and method for tailwater treatment during three periods of the aquaculture of the Litopenaeus vannamei:nursery(0–26 d),middle(27–57 d),and later(57–104 d).The variation of several water parameters during the dissolution of total suspended solid(TSS)in tailwater,applied with the effects of ozone on the microorganism and water quality parameters were investigated.Results showed that the TSS concentrations in tailwater decreased with time,although not significantly(P>0.05),whereas total ammonia nitrogen(TAN),nitrite(NO-2-N),and nitrate(NO_(3)^(-)-N)increased significantly(P<0.05).Therefore,TSS should be removed from the tailwater as early as possible,being most optimal within 4 h.Ozone removed 38.24%–48.95%of TSS,17.78%–90.14%of TAN,and 87.50%–98.90%of NO-2-N after 4 h of treatment.However,it resulted in the significant accumulation of NO_(3)^(-)-N.Moreover,the total number of Vibrio and bacterial counts in aquaculture tailwater was reduced completely by ozone within 4 h.Thus,these results provided technical details and data support for the effective treatment of tailwater from shrimp RAS.

Effects of salinity fluctuation and dietary protein/ carbohydrate ratio on the survival and growth of Juvenile Litopenaeus vannamei

A factorial study was conducted to determine the effects of salinity （constant （SO） and fluctuating （S10）） and isoenergetic diet of different protein/carbohydrate ratios （P/C： 4.1, 1.9, 1.0 and 0.6） on the survival and growth of juvenile Litopenaeus vannamei. The experiment lasted for 35 days. The results showed that I） At S0 and S10, specific growth rate and apparent digestibility coefficient exhibited decreasing trend, while food conversion ratio exhibited increasing trend, with decreasing P/C ratio. At SO, food consumption and specific growth rate of the shrimp fed 4.1 P/C were significantly higher than those fed 1.0 and 0.6 P/C, whereas at S10, no significant differences were detected among different treatments; II） At 4.1 and 1.9 P/C, food consumption and specific growth rate were higher in the shrimp maintained at SO as compared with those at S10, while at 1.0 and 0.6 P/C, an inverse trend was observed. Among four P/C ratios, food conversion ratio of the shrimp at SO was the lowest.

Analysis of Genetic Diversity and Differentiation of Seven Stocks of Litopenaeus vannamei Using Microsatellite Markers

Seven microsatellite markers were used to evaluate the genetic diversity and differentiation of seven stocks of Litopenaeus vannamei, which were introduced from Central and South America to China. All seven microsatellite loci were polymorphic, with polymorphism information content(PIC) values ranging from 0.593 to 0.952. Totally 92 alleles were identified, and the number of alleles(Na) and effective alleles(Ne) varied between 4 and 21 and 2.7 and 14.6, respectively. Observed heterozygosity(Ho) values were lower than the expected heterozygosity(He) values(0.526–0.754), which indicated that the seven stocks possessed a rich genetic diversity. Thirty-seven tests were detected for reasonable significant deviation from Hardy-Weinberg equilibrium. Fis values were positive at five loci, suggesting that there was a relatively high degree of inbreeding within stocks. Pairwise Fst values ranged from 0.0225 to 0.151, and most of the stock pairs were moderately differentiated. Genetic distance and cluster analysis using UPGMA revealed a close genetic relationship of L. vannamei between Pop2 and Pop3. AMOVA indicated that the genetic variation among stocks(11.3%) was much lower than that within stocks(88.7%). Although the seven stocks had a certain degree of genetic differentiation and a rich genetic diversity, there is an increasing risk of decreased performance due to inbreeding in subsequent generations.

Effect of copper on the growth of shrimps Litopenaeus vannamei : water parameters and copper budget in a recirculating system

Shrimps( Litopenaeus vannamei) were intensively cultured in a recirculating aquaculture system for 98 days to investigate effects of 0.3 mg/L Cu on its performance, Cu budget, and Cu distribution. Shrimps in Cu-treated systems had greater mean final weight(11.10 vs 10.50 g), body length(107.23 vs 106.42 mm), survival rate(67.80% vs 66.40%), and yield(6.42 vs 5.99 kg/m 3), and lower feed conversion ratio(1.20 vs 1.29) than those in control systems but the differences were not significant. Vibrio numbers remained stable(10 4 –10 6 colony forming units/mL) in the rearing tanks of both control and treated systems. Total ammonium-N, nitrite-N, nitrate-N, pH, chemical oxygen demand, 5-day carbonaceous biochemical oxygen demand, and total suspended solids were similar in controls and treatments. Dissolved Cu concentration in the treated systems decreased from 0.284 to 0.089 mg/L while in the control systems it increased from 0.006 2 to 0.018 mg/L. The main sources of Cu in the treated systems were the artificially added component(75.7% of total input), shrimp feed(21.0%), water(2.06%), and shrimp biomass(1.22%). The major outputs of Cu occurred via the mechanical filter(41.7%), water renewal(15.6%), and draining of the sediment trap(15.1%). The foam fractionator removed only 0.69% of total Cu input. Harvested shrimp biomass accounted for 11.68% of Cu input. The Cu concentration of shrimps in the Cu-treated systems(30.70 mg/kg wet weight) was significantly higher than that in control systems(22.02 mg/kg). Both were below the maximum permissible concentration(50 mg/kg) for Cu in seafood for human consumption in China. Therefore, recirculating systems can be used for commercial on-growing of Litopenaeus vannamei without loss of shrimp quality, even in water polluted by 0.30 mg/L Cu. The mechanical filter is the main route for Cu removal.

ZResponse to selection,heritability and genetic correlations between body weight and body size in Pacific white shrimp,Litopenaeus vannamei

To quantify the response to selection, heritability and genetic correlations between weight and size ofLitopenaeus vannamei, the body weight （BW）, total length （TL）, body length （BL）, first abdominal segment depth （FASD）, third abdominal segment depth （TASD）, first abdominal segment width （FASW）, and partial carapace length （PCL） of 5-month-old parents and of offspring were measured by calculating seven body measurings of offspring produced by a nested mating design. Seventeen half-sib families and 42 full-sib families ofL. vannamei were produced using artificial fertilization from 2-4 dams by each sire, and measured at around five months post-metamorphosis. The results show that heritabilities among various traits were high： 0.515±0.030 for body weight and 0.394±0.030 for total length. After one generation of selection, the selection response was 10.70% for offspring growth. In the 5th month, the realized heritability for weight was 0.296 for the offspring generation. Genetic correlations between body weight and body size were highly variable. The results indicate that external morphological parameters can be applied during breeder selection for enhancing the growth without sacrificing animals for determining the body size and breed ability; and selective breeding can be improved significantly, simultaneously with increased production.

Molecular cloning and characterization of a threonine/serine protein kinase lvakt from Litopenaeus vannamei

The phosphatidylinositol 3-kinase(PI3K)-AKT pathway is involved in various cellular functions, including anti-apoptosis, protein synthesis, glucose metabolism and cell cycling. However, the role of the PI3K-AKT pathway in crustaceans remains unclear. In the present study, we cloned and characterized the AKT gene lvakt from Litopenaeus vannamei. The 511-residue LVAKT was highly conserved; contained a PH domain, a catalytic domain and a hydrophobic domain; and was highly expressed in the heart and gills of L. vannamei. We found, using Real-Time Quantitative PCR(Q-PCR) analysis, that lvakt was upregulated during early white spot syndrome virus(WSSV) infection. Moreover, the PI3K-specific inhibitor, LY294002, reduced viral gene transcription, implying that the PI3K-AKT pathway might be hijacked by WSSV. Our results therefore suggest that LVAKT may play an important role in the shrimp immune response against WSSV.

Growth and Feed Efficiency of Juvenile Shrimp Litopenaeus vannamei Fed Formulated Diets Containing Different Levels of Poultry by-Product Meal

This feeding trial was conducted to evaluate the potential of poultry by-product meal (PBM) as a protein source in the culture of Litopenaeus vannamei. Seven isonitrogenous and isoenergetic diets were formulated to near to commercial diet with about 40% protein and 7.5% lipid. Fish meal was replaced by 0, 30%, 40%, 50%, 60%, 70% and 100% of PBM (diets 1-7). The diet with 100% fish meal was used as a control (diet 1). Post-larvae were reared in an indoor semi-closed re-circulating system. Each dietary treatment was tested in 4 replicate tanks (260 L) of 40 shrimp, arranged in a completely randomized design. The shrimps were hand-fed for three times a day to near-satiation (0700, 1200 and 1800) for 60 d. Percentage weight gain, survival, feed conversion ratio (FCR), protein efficiency ratio (PER) and body composition of shrimps were measured. There were no significant differences (P>0.05) in growth performance among shrimps fed diets 1-5 (0-60% fish meal replacement). However, shrimps fed diet 7 (100% fish meal replacement) had significantly lower (P<0.05) growth than those fed diets 1-5 (0-60% fish meal replacement). Shrimp fed diets 2-4 (30%-50% fish meal replacement) showed significantly higher growth than those fed diets 6 and 7 (70% and 100% fish meal replacement, respectively). Survival ranged from 94.7% to 100.0% and did not differ significantly (P>0.05) among different experimental diets. No differences in body composition were found among shrimps fed different diets. These results showed that up to 70% of fish meal protein can be replaced by PBM without adversely affecting the growth, survival, FCR, PER and body composition of Litopenaeus vannamei.

Effect of Salinity on the Biosynthesis of Amines in Litopenaeus vannamei and the Expression of Gill Related Ion Transporter Genes

This study examined the effect of salinity on the expression of Na+/K+-ATPase(NKA) α-subunit and vacuolar-type H+-ATPase(V-ATPase) β-subunit gene in the gill of Litopenaeus vannamei. Semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR) assay showed that the expression of NKA α-subunit and V-ATPase β-subunit gene was significantly influenced by salinity. It was found that the NKA activity significantly varied with salinity in time and dose dependent manner; whereas the V-ATPase activity did not. The abundance of NKA α-subunit gene transcript increased rapidly when the salinity decreased from 26 b to 21, and slowly when the salinity decreased from 26 to 31 within the first 24 h. When the salinity decreased from 26 to 21, the transcription of NKA α-subunit gene in gill epithelium was higher at 12 h than that at 0 h, which was consistent with the result of immunoblotting assay of NKA α-subunit. In addition, salinity had a significant time- and dose-dependent effect on the concentration of biogenic amines in both hemolymph and gill. As compared to other parameters, the concentration of dopamine(DA) and 5-hydroxytryptamine(5-HT) varied in different patterns when the salinity decreased from 26 to 21 or increased from 26 to 31, suggesting that DA and 5-HT played different regulatory roles in osmotic adaption and modulation of shrimp when salinity varies.

Regulation of water quality and growth characteristics of indoor raceway culture of Litopenaeus vannamei

Two modes of regulating the water quality of experimental ponds in indoor raceway culture of Litopenaeus vannamei were evaluated using simple water treatment facilities. A self-made water purifying net, aeration stone, composite microbe preparation, and Ceratophyllum demersum were placed in the experimental ponds and the culture water was circulated along the raceway inside the pond using a paddle wheel aerator. In addition, the water quality in the experimental pond was improved by draining effluent from the pipeline at the bottom of ponds 7 and 8 （mode I） and exchanging the circulating water in pond 10 （mode Ⅱ） with the reservoir water in pond 9 using a pump and pipeline. The water quality in the experimental ponds was similar in response to regulation using mode Ⅰ or mode Ⅱ. Water quality parameters in the experimental ponds were controlled within a suitable range by simple facilities during culture period without using any chemical treatments. The rich content of dissolved oxygen was maintained by the circular flow and continuous aeration of the pond water. The respective average values of the main water parameters in experimental ponds 7 and 10 in response to regulation of the water quality using modes Ⅰ and Ⅱwere as follows： pH 8.17 and 7.99; DO 5.16 mg/L and 5.97 mg/L; CODMn18.45 and 12.61 mg/L; TAN （NH3-N） 0.854 mg/L （0.087 mg/L） and 0.427 mg/L （0.012 mg/L）; NO2-N 0.489 mg/L and 0.337 mg/L. Moreover, the average body length and body weight of harvested shrimp of pond 7 and pond l0 were 7.56 cm and 8.99 cm, 5.10 g and 8.33 g, respectively. Furthermore, the survival rate, average biomass yield and average condition factor of the shrimp harvested were 70% and 60%, 2.54 kg/m2 and 2.14 kg/m2, and 0.675 g/cm and 0.927 g/cm, respectively. Linear equations describing the relationship between body length and culture time and cubic or power functions describing the relationship between body weight and body length were obtained based on evaluation of the growth data of shrimps throughout the culture period.

A Study on the Meat and Bone Meal and Poultry By-product Meal as Protein Substitutes of Fish Meal in Practical Diets for Litopenaeus vannamei Juveniles

A study was conducted to evaluate the effects of meat and bone meal (MBM) and poultry by-product meal (PBM) as the replacement of fish meal in the diets on the growth performance, survival and apparent digestibility coefficients (ADC) of Litopenaeus vannamei. The basal diets were formulated with 22% fish meal and other ingredients which provided about 40% protein and 9% lipid in the diet. The experimental diets included MBM or PBM to replace 0, 20%, 40%, 60% and 80% of total fish meal respectively. All diets were iso-nitrogenous and isocaloric in gross terms. The results showed that there were no significant differences (P>0.05) in growth performance and ADC among the treatments fed with the diets in which 0-60% fish meal had been replaced with MBM, while the percent weight gain (WG, %), body length gain (BLG, %) and ADC significantly decreased when the MBM was up to 80% of the fish meal. There were no significant differences (P>0.05) in growth performance and ADC among all the treatments fed with the diets in which 0-80% fish meal had been replaced with PBM.

Characterization of four hemocyanin isoforms in Litopenaeus vannamei

In this study, the gene encoding hemocyanin subunit L, Lv Hc L, was cloned from Litopenaeus vannamei and the genomic organization was characterized. This gene was diverse with many SNPs and also had at least four isoforms, while one of them（Lv Hc L4） only had two exons and the exon2 was missed. Transcription analysis showed that these isoforms of Lv Hc L were up-regulated after WSSV challenge in WSSV-resistant shrimp, while the transcriptions were decreased constantly in WSSV-susceptible shrimp. It is suggested that the hemocyanin had rich polymorphism and was involved in the antiviral response. These results could extend our previous findings and provide insights into the immune feature of hemocyanin, which would be helpful for further studies aimed at antiviral mechanism in invertebrate.

Effect of Replacing Fish Meal with Extruded Soybean Meal on Growth, Feed Utilization and Apparent Nutrient Digestibility of Juvenile White Shrimp(Litopenaeus vannamei)

Extruded soybean meal （ESBM） was evaluated as a protein source for partial replacement of fish meal （FM） in diets of juvenile Litopenaeus vannarnei. In the control diet （Diet 1）, FM protein was replaced with increasing dietary levels of ESBM （4.28%, 8.40%, 12.62%, 16.82%, and 25.26%） at 10%, 20%, 30%, 40%, and 60% levels （Diets 2 to 6, respectively）. An eight-week feeding trial was conducted on 720 juvenile shrimp （0.67 g ± 0.01 g mean initial Weight）, and nutrient digestibility of the six diets was determined. ESBM could replace 20% of FM without causing a significant reduction in growth of shrimp, but other dietary treatments strongly affected whole body composition. Crude protein content of the whole body fed Diet 6 was significantly lower than that fed Diet 2 （P〈0.05）, while crude lipid content of the whole body fed Diet 5 or 6 was significantly higher than that fed Diet 2 （P〈0,05）. Protein digestibilities of Diets 5 and 6 were significantly lower than that of Diet 1 （P〈0.05）. Digestibility of lipids ranged from 96.97% in Diet 6 to 98.34% in Diet 3, whereas dry matter digestibility decreased with increasing replacement level. This study indicates that 20% FM replacement with ESBM in the basic diet containing 40% protein and 30% FM is optimal for juvenile L. vannamei.

Effects of water temperature and dietary carbohydrate levels on growth and energy budget of juvenile Litopenaeus vannamei

A 3×3 factorial experiment was conducted to determine the effects of water temperature (22 ℃, 27℃ and 32℃) and dietary carbohydrate (CBH) levels (15.47%, 29.15% and 41.00%) on growth, food consumption, feed efficiency, apparent digestibility coefficient and energy budget of juvenile Lito- penaeus vannamei. The results showed that, at each dietary CBH level, specific growth rate, food con- sumption and apparent digestibility coefficient generally increased, while feed efficiency decreased with increasing water temperatures. Specific growth rate and food consumption were the highest in the shrimps fed with diet of 29.15% CBH, closely followed by those with 15.47% CBH, and those with 41.00% CBH had the lowest value.

The Effects of Rapid Temperature Changes on HK,PK and HSP70 of Litopenaeus vannamei in Different Seasons

Activities of hexokinase(HK),pyruvate kinase(PK) and levels of HSP70 were measured to evaluate the response of Litopenaeus vannamei to rapid temperature changes under controlled laboratory conditions.Shrimps were subjected to a quick temperature change from 27℃ to 17℃ for the summer case(Cold temperature treatment),or from 17℃ to 27℃ for the winter case(Warm temperature treatment).After 0.5,1,3,6,12,24,48,and 72 h of exposure time,shrimps were sampled and prepared for further analysis.The results showed that the effect of acute temperature changes on activities of HK was significant.Patterns of variations of the two glycolytic enzymes suggested that enzymes in the glycolysis cycle could adjust their activities to meet the acute temperature change.The HSP70 level increased in both cold and warm temperature treatments,suggesting that the rapid temperature changes activated the process of body's self-protection.But the difference in expression peak of HSP70 might be related to the different body size and the higher thermal sensitivity to temperature increase than to temperature decrease of L.vannamei.

Single or combined effects of dietary arabinoxylan-oligosaccharide and inulin on growth performance,gut microbiota,and immune response in Pacific white shrimp Litopenaeus vannamei

The single and synergistic effects of dietary arabinoxylan-oligosaccharide(AXOS)and inulin at different doses(2,4,and 8 mg/g diet)on survival,growth performance,gut microbiota,and immune response in Pacific white shrimp Litopenaeus vannamei were assessed.Singular application of either inulin or AXOS at doses of 4 mg/g diet showed the most stimulatory effects on the growth rate and gene expression levels of chitinase,cathepsin L,chymotrypsin,and extracellular signal-regulated kinase(ERK)in each single prebiotic feeding trial.Compared with single prebiotic treatments,simultaneous application of AXOS and inulin at 4 mg/g diet enhanced remarkably the growth parameters of shrimp and the expression of related genes(chitinase,cathepsin L,chymotrypsin,ERK,myeloid differentiation factor 88,and phenoloxidase)after 8-week feeding(P<0.05).Additionally,gut microbiota analysis indicated the dietary supplementation with combined prebiotics increased significantly the bacterial community richness and relative abundance of Bacillus Pseudomonas Bacteriovorax,and Lactobacillus,and reduced the abundance of Vibrio Rhodococcus,and Photobacterium in the digestive tract of L vannamei.Compared with the single prebiotic treatment and the control,combined prebiotics supplementation boosted notably the survival rate and expression levels of immune-related genes in shrimp infected with Vibrio alginolyticus or white spot syndrome virus.Therefore,simultaneous application of AXOS and inulin shall have a great potential of dietary supplement in the culture of L.vannamei.