摘要
以商品饲料为对照组(A组)、喷涂市售光合细菌菌液(B组)、红假单胞菌菌液(C组)和红螺菌菌液(D组)饲料为试验组,研究饲料喷涂不同光合细菌对凡纳滨对虾幼虾[(1.21±0.24) g]生长、肝胰腺消化酶活性及肠道微生物群落组成的影响。结果显示:光合细菌处理组(B、C和D组)的增重率高于对照组,但差异不显著(P>0.05); C组和D组对虾肝胰腺淀粉酶活力显著高于对照组(P<0.05); B、C和D组对虾肝胰腺脂肪酶活性均高于对照组(P<0.05),肝胰腺蛋白酶活性在各组间无显著性差异(P>0.05)。高通量测序结果表明,随着对虾的生长,其肠道微生物丰富度和多样性均提高。摄食喷涂光合细菌饲料后对虾肠道γ-变形细菌纲所占百分比减少,其中C组和D组γ-变形细菌纲比例显著低于对照组(P<0.05)。各组对虾肠道中检测到的占比大于0.5%的主要微生物属有9个,分别为希瓦氏菌属(Shewanella)、气单胞菌属(Aeromonas)、嗜冷菌属(Algoriphagus)、纤维弧菌属(Cellvibrio)、GpXIII、小梨形菌属(Pirellula)、浮霉状菌属(Planctomyces)、假单胞菌属(Pseudomonas)和红细菌属(Rhodobacter)。使用光合细菌没有显著改变对虾肠道微生物群落的整体丰富度和多样性,但在一定程度上丰富对虾肠道微生物属水平的组成。
In order to evaluate the effect of different dietary photosynthetic bacteria on growth, digestive enzymes and intestinal microbiota of white shrimp Litopenaeus vannamei, four diets(A: control diet; B: Commercial PSB coating diet; C: Rhodopeseudomonas coating diet; and D: Rhodospirillum coating diet) were fed to juvenile shrimp [(1.21±0.24) g]for 2 weeks, respectively. The results showed that: the weight gain rates of the photosynthetic bacteria treatment groups(B, C and D) were slightly higher than that of the control group without significant difference(P>0.05); the amylase activities in hepatopancreas of the shrimp in C and D treatments were significantly higher than that in A treatment(P<0.05); the lipase activities in hepatopancreas of the shrimp in B, C and D treatments were significantly higher than that in A treatment(P<0.05); and there was no significant difference in protease activity among the treatments(P>0.05).The results on sequencing the 16 S RNA genes of microbiota through high-throughput sequencing showed that the diversity and abundance of intestinal microbiota increased along with the shrimp growth. The shrimp fed diets coated with photosynthetic bacteria displayed a reduced proportion on intestinal Gammaproteobacteria accounting for intestinal microflora, the proportion on intestinal Gammaproteobacteria accounting for intestinal microflora of C and D treatments were significantly lower than that of control(P<0.05). Nine main genera(percentage> 0.5%) intestinal microbiota, including Shewanella, Aeromonas, Algoriphagus, Cellvibrio, GpXⅢ, Pirellula, Planctomyces, Pseudomonas and Rhodobacter, coexisted in all the 4 treatments. It therefore suggests that the dietary photosynthetic bacteria application does not significantly alter the overall abundance and diversity of the intestinal microflora, but it enriches the genus composition of the main intestinal flora to a certain extent.
引文
[1] CHAI P C,SONG X L,CHEN G F,et al.Dietary supplementation of probiotic Bacillus PC465 isolated from the gut of Fenneropenaeuschinensis improves the health status and resistance of Litopenaeus vannamei against white spot syndrome virus[J].Fish & Shellfish Immunology,2016,54:602-611.
[2] ADEOYE A A,YOMLA R,JARAMILLO-TORRESA,et al.Combined effects of exogenous enzymes and probiotic on Nile tilapia (Oreochromis niloticus) growth,intestinal morphology and microbiome[J].Aquaculture,2016,463:61-70.
[3] SHA Y J,WANG L,LIU M,et al.Effects of lactic acid bacteria and the corresponding supernatant on the survival,growth performance,immune response and disease resistance of Litopenaeus vannamei[J].Aquaculture,2016,452:28-36.
[4] ZHOU X X,WANG Y B,LI W F.Effect of probiotic on larvae shrimp (Penaeus vannamei) based on water quality,survival rate and digestive enzyme activities[J].Aquaculture,2009,287(3/4):349-353.
[5] LIU H Y,LI Z,TAN B P,et al.Isolation of a putative probiotic strain S12 and its effect on growth performance,non-specific immunity and disease-resistance of white shrimp,Litopenaeus vannamei[J].Fish & Shellfish Immunology,2014,41(2):300-307.
[6] RAMESH D,VINOTHKANNA A,RAI A K,et al.Isolation of potential probiotic Bacillus spp.and assessment of their subcellular components to induce immune responses in Labeorohita against Aeromonashydrophila[J].Fish & Shellfish Immunology,2015,45(2):268-276.
[7] 杨官娥,张肇铭,熊琦.光合细菌化学成分和生理活性研究进展[J].中草药,2005,36(7):1089-1092.YANG G E,ZHANG Z M,XIONG Q.Advances in studies on chemical constituents and physiological activity of photosynthetic bacteria[J].Chinese Traditional and Herbal Drugs,2005,36(7):1089-1092.
[8] 王绍校,杨惠芳,黄志勇,等.嗜盐光合细菌的分离鉴定及其营养成分分析[J].应用与环境生物学报,2003,9(3):298-301.WANG S X,YANG H F,HUANG Z Y,et al.Isolation and identification of marine photosynthesis bacteria and analysis of their nutritional compositions[J].Chinese Journal of Applied and Environmental Biology,2003,9(3):298-301.
[9] 熊晖,梁运祥.外加光合细菌对养殖水体水质及微生物群落的影响[J].湖北农业科学,2011,50(22):4682-4685.XIONG H,LIANG Y X.Study on the impact of external photosynthetic bacteria on water quality and microbial community of aquaculture water[J].Hubei Agricultural Sciences,2011,50(22):4682-4685.
[10] 谢航,陈婕,邱宏端.原生质体融合选育耐高温耐盐红螺菌的研究[J].上海海洋大学学报,2011,20(6):866-872.XIE H,CHEN J,QIU H D.Study on the breeding of Rhodospirillaceae bacteria with salt-resistance and high temperature-resistance by protoplast fusion[J].Journal of Shanghai Ocean University,2011,20(6):866-872.
[11] 王鑫磊,李航,黄旭雄,等.饲料中胆固醇含量对淡水养殖凡纳滨对虾生长性能、抗弧菌和抗亚硝态氮胁迫能力的影响[J].动物营养学报,2016,28(8):2639-2649.WANG X L,LI H,HUANG X X,et al.Effects of dietary cholesterolcontent on growth performance,Vibrio and nitrite nitrogen stress resistant abilities of Litopenaeus vannamei cultured in freshwater[J].Chinese Journal of Animal Nutrition,2016,28(8):2639-2649.
[12] 刘双印,徐龙琴,李道亮,等.基于物联网的南美白对虾疾病远程智能诊断系统[J].中国农业大学学报,2014,19(2):189-195.LIU S Y,XU LQ,LI D L,et al.Research on remote system for disease diagnosis of Penaeus vannamei based on Internet of things[J].Journal of China Agricultural University,2014,19(2):189-195.
[13] GARFIAS-LóPEZ J A,CASTRO-ESCARPULI G,CáRDENASP E,et al.Immunization with intestinal microbiota-derived Staphylococcus aureus and Escherichia coli reduces bacteria-specific recolonization of the intestinal tract[J].Immunology Letters,2018,196:149-154.
[14] CHEN GJ,XIE MH,WAN P,et al.Digestion under saliva,simulated gastric and small intestinal conditions and fermentation invitro by human intestinal microbiota of polysaccharides from Fuzhuan brick tea[J].Food Chemistry,2018,244:331-339.
[15] CHUMPOL S,KANTACHOTE D,NITODA T,et al.The roles of probiotic purple nonsulfur bacteria to control water quality and prevent acute hepatopancreatic necrosis disease (AHPND) for enhancement growth with higher survival in white shrimp (Litopenaeus vannamei) during cultivation[J].Aquaculture,2017,473:327-336.
[16] 王梦亮,郭小青,梁生康,等.光合细菌(PSB)对鲤鱼肠道菌群及肠消化功能的影响[J].中国微生态学杂志,1999,11(3):146-148.WANG M L,GUO X Q,LIANG S K,et al.Effect of photosynthetic bacteria on intestinal microflora and the activity of digestive enzyme of carps[J].Chinese Journal of Microecology,1999,11(3):146-148.
[17] 陆文清,李德发,任继平,等.福林显色法测定饲料用蛋白酶活力的要点分析[J].饲料研究,2003(9):29-30.LU W Q,LI D F,REN J P,et al.Key analysis of the determination of protease activity in feed by Folin chromatography[J].Feed Research,2003(9):29-30.
[18] 李蓓,李晓晖,衣杰荣.对硝基苯酚法对雅致放射毛霉脂肪酶特性的研究[J].食品工业科技,2011,32(11):220-222,263.LI B,LI X H,YI J S.Determination on lipase enzymatic characteristics and activity of Actinomucorelegans by pNP method[J].Science and Technology of Food Industry,2011,32(11):220-222,263.
[19] CAO Y C,WEN G L,LI Z J,et al.Effects of dominant microalgae species and bacterial quantity on shrimp production in the final culture season[J].Journal of Applied Phycology,2014,26(4):1749-1757.
[20] 王刚,朱站英,华雪铭,等.国产鱼粉饲料中添加卵黄抗体、丁酸钠和枯草芽孢杆菌对中华鳖生长和摄食偏好的影响[J].上海海洋大学学报,2017,26(3):376-382.WANG G,ZHU Z Y,HUA X M,et al.Effect of egg yolk antibody,sodium butyrate and Bacillus subtilis supplementation in domestic fish meal based feed on growth and feeding preference of China soft-shelled turtle[J].Journal of Shanghai Ocean University,2017,26(3):376-382.
[21] 李卓佳,林亮,杨莺莺,等.芽孢杆菌制剂对凡纳滨对虾Litopenaeus vannamei肠道微生物群落的影响[J].南方水产,2005,1(3):54-59.LI Z J,LIN L,YANG Y Y,et al.Effect of Bacillus commercial probiotic on intestinal microflora of white shrimp,Litopenaeus vannamei[J].South China Fisheries Science,2005,1(3):54-59.
[22] DUAN Y F,ZHANG Y,DONG H B,et al.Effect of dietary Clostridium butyricum on growth,intestine health status and resistance to ammonia stress in Pacific white shrimp Litopenaeus vannamei[J].Fish &Shellfish Immunology,2017,65:25-33.
[23] 唐杨,刘文亮,宋晓玲,等.饲料中补充蜡样芽孢杆菌对凡纳滨对虾生长及其肠道微生物组成的影响[J].水产学报,2017,41(5):766-774.TANG Y,LIU W L,SONG X L,et al.Effects of dietary with Bacillus cereus on the growth rate and intestinal microflora of Litopenaeus vannamei[J].Journal of Fisheries of China,2017,41(5):766-774.
[24] WANG Y B,XU Z R.Effect of probiotics for common carp (Cyprinus carpio) based on growth performance and digestive enzyme activities[J].Animal Feed Science and Technology,2006,127(3/4):283-292.
[25] WANG Y B.Effect of probiotics on growth performance and digestive enzyme activity of the shrimp Penaeus vannamei[J].Aquaculture,2007,269(1/4):259-264.
[26] REUTER J A,SPACEK D V,SNYDER M P.High-throughput sequencing technologies[J].Molecular Cell,2015,58(4):586-597.
[27] 李存玉,徐永江,柳学周,等.池塘和工厂化养殖牙鲆肠道菌群结构的比较分析[J].水产学报,2015,39(2):245-255.LI C Y,XU Y J,LIU X Z,et al.Comparative analysis of composition,diversity and origin of intestinal bacterial community in pond-and indoor tank-culture Japanese flounder (Paralichthys olivaceus)[J].Journal of Fisheries of China,2015,39(2):245-255.
[28] 查广才,周昌清,麦雄伟.凡纳滨对虾低盐度高产虾池环境微生物生态研究[J].生态科学,2006,25(3):231-235.ZHA G C,ZHOU C Q,MAI X W.Environmental bacteria in Litopenaeus vannamei low salinity intensive stocking ponds[J].Ecologic Science,2006,25(3):231-235.
[29] YANG W,ZHENG C,ZHENG Z M,et al.Nutrient enrichment during shrimp cultivation alters bacterioplankton assemblies and destroys community stability[J].Ecotoxicology and Environmental Safety,2018,156:366-374.
[30] ZHENG Y F,YU M,LIU Y,et al.Comparison of cultivable bacterial communities associated with Pacific white shrimp (Litopenaeus vannamei) larvae at different health statuses and growth stages[J].Aquaculture,2016,451:163-169.
[31] ZHANG M L,SUN Y H,CHEN K,et al.Characterization of the intestinal microbiota in Pacific white shrimp,Litopenaeus vannamei,fed diets with different lipid sources[J].Aquaculture,2014,434:449-455.
[32] DEVARAJA T N,YUSOFF F M,SHARIFF M.Changes in bacterial populations and shrimp production in ponds treated with commercial microbial products[J].Aquaculture,2002,206(3/4):245-256.
[33] HOSTINS B,LARA G,DECAMP O,et al.Efficacy and variations in bacterial density in the gut of Litopenaeus vannamei reared in a BFT system and in clear water supplemented with a commercial probiotic mixture[J].Aquaculture,2017,480:58-64.
[34] 闵涛玲,杨启银,曩潇潇,等.光合细菌菌剂中-株粘质沙雷氏菌的分离与鉴定[J].安徽农业科学,2006,34(22):5837-5838,5841.MIN T L,YANG Q Y,NANG X X,et al.Isolation and identification of a Serratiamarcescens from photosynthetic bacteria[J].Journal of Anhui Agricultural Sciences,2006,34(22):5837-5838,5841.
[35] MIZUNO Y,NARIMATSU H,KISHI Y,et al.Structural problems of medical news reports in newspapers:a verification of news reports on an incident of mass nosocomial Serratia infection[J].Journal of Infection and Chemotherapy,2010,16(2):107-112.