七种养殖特色淡水鱼脏器系数的测定和比较

为研究养殖特色淡水鱼成鱼主要脏器系数,并探讨各品种组织器官重量与体重的关系,本研究对杂交鲟(俄罗斯鲟Acipenser gueldenstaedti♀×史氏鲟A.schrenckii♂)、斑点叉尾■(Ietalurus punetaus)、鳜(Siniperca chuatsi)、大口黑鲈(Micropterus salmoides)、黄颡鱼(Pelteobagrus fulvidraco)、黄鳝(Monopterus albus)和泥鳅(Misgurnus anguillicaudatu)7种特色淡水鱼主要内脏器官测定和比较,并对各脏器重量和体重进行相关性分析,拟合回归方程。结果显示,除黄鳝鳃退化无此器官系数外,7种特色淡水鱼的脏器系数中心脏系数最小,肾脏、胃和肠的脏器系数居中,肝脏系数在所测器官中最大。脏器与体重关系上,杂交鲟肝脏和肠,斑点叉尾■肾脏、脾脏、胃、心脏、鳃、胆囊和肝脏,鳜肠和心脏,大口黑鲈鳃、眼睛、肝脏、脾脏、肠、胃,黄颡鱼鳃、眼睛、肾脏、胃、肠、心脏和肝脏,黄鳝胆囊、肝脏、胃、肾脏和肠,泥鳅鳃、眼睛、脾脏和肠的重量与体重呈显著正相关,符合线性关系。不同性别上,鳜的肝脏系数和大口黑鲈的鳃系数雌、雄之间差异显著,其他脏器系数无显著差异。结果表明,不同品种鱼类之间脏器系数存在显著差异,不同鱼类主要脏器重量与体重之间普遍存在一定线性关系。     

大口黑鲈leptin A基因的组织表达及其对急性低氧胁迫的响应

瘦素(leptin)是一种重要的激素,参与调节动物的摄食、繁殖和能量消耗,其可以通过抑制食欲和促进能量代谢的方式维持机体能量稳态。大多数鱼类具有双重瘦素基因。在本研究中,利用PCR技术克隆了大口黑鲈leptin A基因的编码区,其开放阅读框(ORF)为486 bp,编码161个氨基酸蛋白。通过与其他物种进行同源性比对,发现鲈形目的 leptin基因的保守性较高,一致性高达91.46%,在大口黑鲈leptin A中几乎不存在基因特异性突变位点,而且大口黑鲈leptin A氨基酸序列与鳜同源性最高(92.59%),与花鲈(89.51%)、布氏鲳鲹(87.04%)、斜带石斑鱼(83.87%)的同源性次之。组织分布结果显示,leptin A基因在肝脏中的表达量最高,在心脏、头肾、脑、中肾、肠、脾脏、鳃、肌肉和性腺中的表达量均较低。此外,对大口黑鲈进行不同程度急性低氧胁迫[重度低氧(1.2±0.2)mg/L和中度低氧(3.5±0.2)mg/L],发现低氧胁迫初期时,严重低氧组和中度低氧组的leptin A的表达量都升高,显著高于对照组Leptin A的表达。这表明急性低氧能够诱导大口黑鲈leptin A在肝脏中的表达。综上所述,大口黑鲈leptin A基因与鳜leptin A基因的同源性最高,leptin A主要在大口黑鲈肝脏中显著表达,急性低氧胁迫能显著诱导leptin A的表达。     

大口黑鲈两种脂蛋白脂肪酶基因cDNA的克隆及表达特征分析

为促进肉食性鱼类人工配合饲料开发的理论基础研究,分析鱼类脂肪代谢的机制,实验克隆了大口黑鲈2个脂蛋白脂肪酶基因LPLtype1和LPLtype2的cDNA。序列分析表明,LPLtype1基因cDNA序列全长2 156 bp,编码516个氨基酸;LPLtype2基因cDNA序列全长1 710 bp,编码346个氨基酸。大口黑鲈LPLtype2与LPLtype1氨基酸序列之间的同源性为43.5%。系统进化分析表明,大口黑鲈LPLtype1和鳜LPL聚为一支,大口黑鲈LPLtype2和大麻哈鱼LPLtype2紧密聚为一支。预测分析发现,大口黑鲈LPLtype1和LPLtype2基因编码蛋白的活性中心位点、N-糖基化位点、二聚体形成的保守疏水残基位点、肝素结合域等主要功能域与硬骨鱼类和其他脊椎动物对比都比较保守。运用实时定量PCR方法检测脂蛋白脂肪酶mRNA的组织分布,发现LPLtype1和LPLtype2都在肝脏中表达量最高,推测这与肝脏是最主要的营养诱导性储脂部位有关。     

草鱼碱性氨基酸转运载体y+LAT2 cDNA基因克隆与表达研究

利用逆转录聚合酶链式反应(RT-PCR)和cDNA末端快速扩增(RACE)方法,克隆y+LAT2基因cDNA序列,并利用实时荧光定量PCR探讨y+LAT2在草鱼(Ctenopharyngodon idellus)各组织中的表达情况以及饥饿对其mRNA的影响。结果表明,获得的y+LAT2基因的cDNA序列片段为1849bp,含1371bp的核心序列,编码456个氨基酸。预测草鱼氨基酸序列与斑马鱼(Danio rerio)的同源性高达96.5%,与哺乳动物和两栖动物的同源性在78%～83%,构建的系统进化树与传统形态学分类一致。在草鱼的肌肉、脑、鳃、心、肾脏、肝、后肠、中肠、前肠、脾脏组织均检测到y+LAT2基因的表达。草鱼脾脏、肾脏以及前肠、中肠的y+LAT2 mRNA表达水平对禁食的反应不同,在短期饥饿(14d)期间,其y+LAT2 mRNA表达水平均呈下降趋势。草鱼碱性氨基酸转运载体y+LAT2基因全长cDNA序列的获得,有利于进一步探讨鱼类氨基酸的吸收转运机制。     

急性高温胁迫对大口黑鲈“优鲈3号”组织损伤及HSPs基因表达的影响

将25℃(对照组)下暂养的体质量(9.87±1.20) g大口黑鲈"优鲈3号"幼鱼直接放入28、31、34、37℃水环境中,0、6、48 h时取肝脏和鳃组织,探究不同温度应激后对大口黑鲈"优鲈3号"幼鱼肝脏、鳃组织结构损伤及HSPs基因在这些组织中表达的影响。试验结果显示,28℃和31℃组肝脏和鳃组织均与对照组无差异,34℃肝脏和鳃组织出现轻微的细胞水肿,37℃则出现肝细胞空泡化、细胞间界限杂乱模糊、细胞核溶解,鳃小片末端出现膨大且弯曲,部分鳃小片因红细胞过多而涨破。在不同水温下,HSPs基因的表达量变化不同。6 h时,肝脏和鳃组织中HSP70和HSC70 mRNA表达量均随着胁迫温度的升高而升高,在37℃时表达量最高;48 h时,肝脏和鳃组织中HSP70和HSC70 mRNA表达量相较于6 h均出现不同程度的下降(除31℃组鳃组织HSP70);且随着胁迫温度的升高呈先升后降的趋势,37℃组的表达量最低。结果表明,高温胁迫会使大口黑鲈"优鲈3号"幼鱼产生一定应激损伤,因此在实际养殖生产中,应密切关注温度的变化,降低温度胁迫对鱼体免疫机能的影响。     

10种免疫相关基因在斜带石斑鱼组织中的表达分析

采用相对实时荧光定量PCR(Quantitative Real-time PCR,qRT-PCR)技术,以β-肌动蛋白(β-actin)的表达量作为内参,对健康养殖斜带石斑鱼(Epinephelus coioides)不同组织(头肾、心脏、肝脏、脾脏、鳃、肌肉、鳍、眼、肠道)中的10种免疫相关基因:免疫球蛋白(IgM)、CD4、MHCIIa、TCRβ、CD8a、MHCIa、ISP16、Mx、TNFR14、HSP90的mRNA表达量进行了研究。结果显示,10种免疫相关基因在斜带石斑鱼的10种组织中均有表达。其中体液免疫相关基因[免疫球蛋白(IgM)、CD4和MHCIIa]在鳃中的表达量最高,在其他组织部位的表达量相对较少。细胞免疫相关基因(TCRβ、CD8a和MHCIa)也是在鳃中的表达量最高,在眼、肌肉、肝和肠的部位表达量相对较少。ISP16在鳃和肝中的表达量较高,在肌肉和肠中的表达量较低。Mx在鳃中的表达量较高,在肌肉和肠的表达量较低。TNFR14在眼的表达量较高,在肠的表达量较低。HSP90在鳃和肝中的表达量相对较高,在心和肌肉的表达量较低。     

大口黑鲈蛙病毒分子流行病学及组织病理分析

为探讨大口黑鲈蛙病毒（LMBV）的分子流行规律及在感染后的组织病理变化，实验对2019—2021年采集的723份患病大口黑鲈样品进行荧光定量PCR （qPCR）检测。结果显示，LMBV的阳性率为63.62%，挑选阳性样品接种鳜脑细胞（Chinese perch brain cells,CPB），共获得93株LMBV。通过对分离株MCP、ATP酶、DNA聚合酶和甲基转移酶基因进行PCR扩增与测序分析，发现上述基因在LMBV分离株中均保守，其中MCP、ATP酶基因一致性均为100.0%、甲基转移酶基因一致性为99.7%～100.0%、DNA聚合酶基因一致性为99.8%～100.0%。选取1株LMBV毒株感染健康大口黑鲈，采用qPCR方法对不同组织中的病毒载量进行检测，结果显示，大口黑鲈蛙病毒在心脏、肝脏、脾脏、肾脏、胃、肠和脑等组织中均有分布，人工感染LMBV后的前5天病毒载量逐步升高；感染后第5天，心脏中病毒载量最高（9.5×10⁵个/mg拷贝），脑组织中病毒载量最低（2.9×10²个/mg拷贝）。组织病理结果表明，LMBV可导致大口黑鲈多种组...     

草鱼HSP90基因cDNA序列克隆及其组织表达差异

根据斑马鱼HSP90(Heat shock protein)序列(NM_131328)设计并合成一对引物,提取草鱼肝脏组织总RNA,RT-PCR扩增获得草鱼HSP90基因cDNA部分序列596 bp,获得GenBank登陆号为FJ517554.将测序结果利用DNAman软件与其他几种已知序列的鱼进行比对,结果表明,草鱼HSP90与斑马鱼、鲤、鲋的同源性依次为90%、89%、92%.同时利用半定量(Semi-quantitative,SQ)RT-PCR技术检测HSP90在草鱼脂肪、肌肉、肠、脑、粘液、性腺、鳔、肝脏、心脏、脾脏、鳃、鳍12个组织的表达差异.结果表明,HSP90在草鱼除了鳍外的其他11个组织中均检测到表达,其中在心脏中表达最高,但与鳃、性腺、脑、脾脏中的表达无显著差异(P>0.05),在脂肪、粘液、鳔组织中的表达显著低于其他几种组织(P<0.05),在肝脏、肌肉与肠中的表达无显著差异(P>0.05).     

禁食对大口黑鲈生长和肝脏IGF-I mRNA表达丰度的影响

运用实时荧光定量PCR技术,研究禁食对体质量为(100+1)g的1龄大口黑鲈(Micropterus salmoides)的生长和肝脏中类胰岛素生长因子-I(IGF-I)mRNA表达丰度的影响.在为期6周的实验期间,对照组每天表观饱食投喂2次;禁食组禁食3周后恢复投喂3周.实验期间对照组鱼体质量和体长逐渐增加.6周后,对照组鱼体增重率为10.99%,体长增加率为3.07%.禁食组禁食3周期间,鱼体质量下降了5.08%、体长减少了1.79%.同时肝组织IGF-I mRNA表达水平呈下降趋势,禁食结束时仪为对照组的29.93%.恢复投喂2周后肝组织IGF-I mRNA的表达量仍显著低于对照组(P＜0.05),鱼体质量为实验初始水平的98.54%,体长为实验初始水平的99.03%;恢复投喂3周后,禁食组鱼体质量、体长恢复到实验初始时水平(P＞0.05),但仍与对照组有显著差异(P＜0.01);肝组织IGF-I mRNA的表达丰度与对照组无显著性差异(P＞0.05).结果显示,禁食使大口黑鲈体质量、体长下降,同时肝组织IGF-I mRNA表达丰度也随之降低;而恢复投饲后,其生长、肝脏IGF-I mRNA的表达丰度也逐渐恢复.研究表明,鱼类的营养状况、生长和与IGF-I mRNA的表达之间存有正相关关系.     

草鱼孕烷X受体与细胞色素P450 3A mRNA表达相关性初步研究

根据GenBank中斑马鱼(Danio rerio)和虹鳟(Oncorhynchus mykiss)孕烷X受体(pregnane X receptor,PXR)基因序列设计保守区域简并引物,通过PCR扩增获得草鱼(Ctenopharyngodon idellus)PXR cDNA核苷酸序列片段为509 bp,经推导获取169 aa序列。对草鱼与其它物种PXR氨基酸序列进行同源性比较,用以鉴定其在物种中的进化。应用实时荧光定量PCR(qRT-PCR)检测草鱼9种组织PXR和细胞色素P450 3A(cytochrome P450 3A,CYP3A)基因mRNA相对含量,结果表明,其表达丰度依次为:前肠﹥肝脏﹥肾脏﹥鳃﹥肌肉﹥后肠﹥性腺﹥心脏﹥脾脏,CYP3A和PXR基因在草鱼组织中表达分布基本一致,在前肠、肝脏和肾脏高度表达,而在其它组织低度表达。为了进一步研究PXR和CYP3A基因表达相关性,通过细胞,特选用诱导剂利福平(rifampicin,RIF),最佳诱导浓度40μM,孵育1、2、4、6、8、10、12、24 h后,用qRT-PCR检测CYP3A-PXR基因动态表达过程。结果显示,诱导组CYP3A和PXR基因表达量均高于对照组,显示出显著诱导效应。     

Effects of different dietary lipids on growth,body composition and lipid metabolism‐related enzymes and genes in juvenile largemouth bass,Micropterus salmoides

This study investigates the effects of different lipids on growth, body composition and lipid metabolism of largemouth sea bass fish Micropterus salmoides. A total of 360 juvenile M. salmoides (mean ± SD mass = 33.83 ± 0.15 g) were randomly stocked into 12 tanks of 0.5 m³ volume for 8 weeks. Four replicates were made in each group, which were fed one of three diets containing fish oil (FO), soybean oil (SO) or lard oil (LO). The weight gain rate and specific growth rate did not differ among the groups (p > 0.05). Fish oil fish had the lowest condition factor (p < 0.05) and highest serum glucose content (p < 0.05). Crude lipid contents in the whole body and in the liver and muscle of FO fish were significantly lower than in the SO and LO groups (p < 0.05). The fatty acid composition of whole‐body lipids was closely correlated with that of the diet. The carnitine palmitoyltransferase 1 (cpt1) activity in the FO group was significantly higher than those in the SO and LO groups (p < 0.05). No significant differences in fatty acid synthase (fasn) activity were observed among the groups (p > 0.05). The Cpt1 and fasn gene expression levels in the FO group were significantly higher than those of the SO and LO groups (p < 0.05). The apolipoprotein B100 gene expression level was significantly higher in the SO group than in the FO group (p < 0.05). Fatty acid‐binding protein 1 gene expression levels in the FO and SO groups were not different (p > 0.05) but were both higher than that of the LO group (p < 0.05). The delta‐6 fatty acyl desaturase gene expression level in the LO group was significantly higher than that in the FO group (p < 0.05), but lower than that in the SO group (p < 0.05). It can be concluded that FO can be completely replaced by SO or LO in the M. salmoides diet, at least within the 8‐week culture period. Different types of dietary lipids significantly affect body condition and hepatic lipid metabolism in M. salmoides.     

High dietary starch impaired growth performance,liver histology and hepatic glucose metabolism of juvenile largemouth bass,Micropterus salmoides

A 60‐day experiment was carried out to investigate dietary starch levels on growth performance, hepatic glucose metabolism and liver histology of largemouth bass, Micropterus salmoides. Fish (initial weight 22.00 ± 0.02 g) were fed five graded levels of dietary corn starch (0, 50, 100, 150 and 200 g/kg). Fish fed low (0 and 50 g/kg) dietary starch showed significantly higher weight gain than other groups (p < .05). Liver lipid and glycogen accumulations were induced when dietary starch higher than 100 g/kg. After 20 days of feeding, hexokinase activity and mRNA expression were decreased in fish fed dietary starch higher than 150 g/kg (p < .05) and the pyruvate kinase showed the opposite tendency. Insulin receptor 1 (irs1), glucagon‐like peptide‐1 receptor and glucose transport protein 2 (glut2) mRNA expression were decreased with the increasing dietary starch after 10 days of feeding (p < .05). These results indicated gluconeogenesis was depressed and β‐oxidation was enhanced in response to high dietary starch, while the glycolysis was inhibited and endocrine system was impaired when fish fed high dietary starch; then, glucose homeostasis was disturbed and finally led to the glucose intolerance of largemouth bass.     

Effect of dietary oxidized fish oil on growth performance,body composition,antioxidant defence mechanism and liver histology of juvenile largemouth bass Micropterus salmoides

Four isonitrogenous and isolipidic diets containing fresh fish oil (peroxide value, POV: 11.5 meq kg^?1, diet FR) and three degrees of oxidized fish oil (POV: 132, 277 and 555 meq kg^?1, diet OX₁₃₂, OX₂₇₇ and OX₅₅₅, respectively) were formulated to investigate the effects of dietary oxidized fish oil on growth performance, body composition, antioxidant defence mechanism and liver histology of juvenile largemouth bass. After a 12‐week feeding trail, a proportion of approximately 9% of Micropterus salmoides showed inflammation and haemorrhage at the base of dorsal, pectoral and tail fin in both groups OX₂₇₇ and OX₅₅₅. Fish fed oxidized oil diets obtained significantly higher (P < 0.05) weight gain and specific growth rate because of their remarkable higher feed intakes, compared with the fresh oil receiving group. The analysis of biometric parameters and body composition indicated significant differences (P < 0.05) in various test diets. The activities of hepatic catalase and superoxide dismutase were significantly stimulated (P < 0.05) by oxidized oil ingestion. Hepatic glutathione peroxidase, glutathione reductase and glutathione‐S‐transferase activities were significantly higher (P < 0.05), and liver glutathione content was markedly lower (P < 0.05) in group OX₅₅₅ than the other treatments. Oxidized oil consumption resulted in marked depletion (P < 0.05) of vitamin E concentration in plasma, liver and muscle tissue, increased plasma and muscle malondialdehyde content along with decreased haematocrit value. Histological examinations indicated that hepatocytes with lipid vacuoles and nuclear migration were shown in groups OX₂₇₇ and OX₅₅₅. The overall results in this study suggested that an increased oxidative stress in M. salmoides fed oxidized lipid may account for their stimulated hepatic antioxidant defences, vitamin E depletion in plasma and certain tissues, and pathological changes. The detrimental effect of oxidation products on fish health and the unexpectedly enhanced feed intake of oxidized feeds in M. salmoides underline the importance that cares should be taken to minimize dietary oxidation products to the greatest extent possible.     

大口黑鲈致死性结节病病原的分离、鉴定及组织病理学观察

2018年4月四川邛崃地区某养殖场大口黑鲈感染致死性结节病,死亡率高达80%。为明确病因,通过常规微生物分离、鉴定,从患病鱼皮肤溃疡灶、鱼鳔腔积液和肝脏组织中分离到同一株优势菌,命名为HSY-NS02。经菌体形态观察、革兰氏染色和抗酸染色镜检、生理生化实验、16S rDNA序列扩增及系统发育分析和特异性PCR扩增,确定该菌为诺卡氏菌。进一步对健康大口黑鲈进行分离菌的人工感染实验以确定其致病性,结果显示,人工感染鱼出现与自然发病相似症状,且从人工感染鱼体中再次分离到相同菌。组织病理学观察显示,皮肤溃疡灶、心脏、肝脏、脾脏、肾脏和鳃发生不同程度的慢性炎性肉芽肿病变,其中脾脏病变最为严重。对菌株HSY-NS02进行药物敏感性实验,结果显示,该菌对庆大霉素、新霉素和制霉菌素3种药物敏感,对其他18种药物耐受,呈多重耐药性。     

Molecular cloning,characterization and expression analysis of glucose transporters from Rachycentron canadum

It is very necessary to clarify the mechanism of carbohydrate metabolism in fish due to their poor ability to utilize carbohydrates. Glucose transporters (GLUTs) are important carriers involved in glucose transport across the plasma membrane, which is the first rate‐limiting step of carbohydrate metabolism. In this study, five glucose transporters (RcGLUT1, RcGLUT2, RcGLUT3, RcGLUT5 and RcGLUT9) were obtained from Rachycentron canadum. The complete cDNAs open reading frames (ORF) of RcGLUT1, RcGLUT2, RcGLUT3, RcGLUT5 and RcGLUT9 were 1,476 bp, 1,530 bp, 1,551 bp, 1,539 bp and 1,578 bp (GenBank accession no. MK560257 , MK560258 , MH184460 , MH184461 and MH184462 ), encoding 491, 509, 516, 512 and 525 amino acid (aa) residues respectively. All five RcGLUTs contained a Sugar_tr domain, which is an important feature of the GLUT gene family. The multiple sequence alignment and phylogenetic relationship analyses showed that RcGLUTs were highly conserved and homologous from fish to mammals. Examination of tissue distribution showed that RcGLUTs were expressed constitutively in R. canadum. RcGLUT1‐5 and RcGLUT9 had the highest expression in the foregut, kidneys, haemocytes, muscles, liver and heart respectively. All six RcGLUTs first increased to peak levels of expression and then reduced both in the liver and muscle after fasting. The exception is that the expression levels of RcGLUT4 and RcGLUT5 in muscle were significantly lower than the control. In response to hypoxia, only RcGLUT2 in the liver and muscle and RcGLUT9 in muscle were expressed at a significantly lower level relative to the control. In sum, all RcGLUTs in the liver and muscles showed significant changes in response to fasting and hypoxia, suggesting that GLUT genes may play a role in the response to common physiological stresses.     

大口黑鲈鲁氏耶尔森氏菌的分离鉴定、主要毒力基因及致病性研究

为探明2021年3月福建某水库养殖大口黑鲈疾病暴发的原因,实验从患病大口黑鲈肝脏、肾脏、脾脏中分离优势菌,通过人工感染实验确定病原菌,并综合16S rDNA基因序列分析、生理生化和质谱特征等技术对该病原菌进行种属鉴定,同时,进行毒力基因检测、药物敏感性实验以及组织病理学观察。结果显示,从病鱼脾脏中分离获得1株优势菌并鉴定为鲁氏耶尔森氏菌;该菌株对大口黑鲈的半致死剂量为3.8×10⁵ CFU/尾;该菌株携带yrP1、yhl A、yhl B等毒力基因,对恩诺沙星、盐酸多西环素、氟甲喹、硫酸新霉素、氟苯尼考等5种药物相对敏感。组织病理学观察发现,鲁氏耶尔森氏菌的感染造成大口黑鲈肝脏、肾脏、脾脏不同程度的损伤,表现为明显的变性、坏死及炎症细胞的浸润等。本研究首次报道了鲁氏耶尔森氏菌对养殖大口黑鲈的致病性,可为养殖大口黑鲈鲁氏耶尔森氏菌病的诊断和药物防治提供参考依据。     

阿克曼菌对鲤糖代谢的调控作用研究

[目的]本试验旨在探究巴氏灭活的阿克曼菌（Pasteurized Akkermansia muciniphila，P-Akk）调控鲤（Cyprinus carpio L.）糖代谢的分子机制。[方法]本研究通过不同糖水平（20%、30%、40%、50%葡萄糖）试验，探究鲤（10.5±1 g）4周和8周时胰高血糖素样肽-1（glucagon-like peptide-1，GLP-1）分泌及阿克曼菌（Akkermansia muciniphila，Akk）定植的时空变化；在不同糖水平试验基础上，设置40%葡萄糖和三个不同浓度P-Akk（108 cfu/g P-Akk、109 cfu/g P-Akk、1010 cfu/g P-Akk）组，探究添加P-Akk 4周后对鲤（16.38±0.39 g）糖代谢及GLP-1的调控；通过P-Akk孵育鲤原代肝、肠细胞试验，共同探究不同糖水平下鲤肠道Akk、GLP-1对糖代谢的调控机制。[结果]结果显示：（1）随着饲料中葡萄糖含量的升高，鲤血清中GLP-1b含量显著降低（P < 0.05），4周时高糖组鲤肠道GLP-1a、GLP-1b以及肠道内容物中Akk丰度均显著降低（P < 0.05），但8周时无显著性差异（P > 0.05）。（2）P-Akk处理后，鲤血清中葡萄糖、GLP-1含量显著减少（P < 0.05）；中肠绒毛高度、肌层厚度显著增加，GLP-1含量减少，短链脂肪酸含量增加，muc2 mRNA表达水平升高（P < 0.05）；此外，P-Akk上调肝脏中ampk、pi3k、akt及糖酵解基因gk、pfk mRNA表达量，下调糖异生基因pepck mRNA表达量（P < 0.05）。（3）原代肝、肠细胞孵育结果显示，P-Akk处理肠细胞后GLP-1含量显著减少（P < 0.05），而gpr40的mRNA表达量升高；此外肝细胞中糖酵解基因gk、pfk mRNA表达量显著升高。[结论]综上，外源添加P-Akk可以缓解高糖饲料引起的血糖升高，维持葡萄糖稳态。[意义]该研究结果可为Akk作为益生菌在水产饲料中的应用提供理论基础。     

Triploidy Induction in Largemouth Bass,Micropterus salmoides

Triploid largemouth bass, Micropterus salmoides, were produced by using hydrostatic pressure treatments five minutes after fertilization. Treatments ranged from 4,000 p.s.i. for 3 minutes to 8,000 p.s.i. for 1 minute was best, yielding 100% triploids and a relatively low mortality.