酵母水解物对吉富罗非鱼生长、免疫力、抗氧化性及抗无乳链球菌感染的影响

本研究旨在探究饲料中添加酵母水解物对吉富罗非鱼(Oreochromis niloticus)生长性能、免疫力和抗病力的影响。在基础饲料中添加0 (对照组)、0.5%、1.0%、1.5%和2.0%的酵母水解物制成5种实验饲料,对5组初始体重为(25.80±0.45) g的吉富罗非鱼进行为期8周的养殖实验,每组 3个重复,每个重复30尾鱼。实验结束后,每个重复选择12尾鱼进行无乳链球菌(Streptococcus agalactiae)攻毒实验,每尾鱼腹腔注射0.2 mL浓度为8.3×105 CFU/mL的菌液,连续观察14 d,统计累积死亡率。结果显示,随着酵母水解物添加量的增加,吉富罗非鱼终末体质量(FBW)和特定生长率(SGR)均呈先显著上升(P<0.05)后下降的趋势。回归分析表明,当SGR达到最大值时,对应的酵母水解物添加量为1.29%。罗非鱼血清一氧化氮合酶、酚氧化酶、酸性磷酸酶、过氧化氢酶和超氧化物歧化酶活性均随饲料中酵母水解物添加量的提高呈先上升后下降的趋势(P<0.05),添加量为1.5%时达最大值。饲料添加酵母水解物对吉富罗非鱼无乳链球菌的抵抗力有显著影响(P<0.05),除添加量为0.5%的实验组死亡率与对照组差异不显著外,其他实验组罗非鱼的死亡率均显著低于对照组(P<0.05)。本研究中,饲料添加酵母水解物可有效提高吉富罗非鱼的生长性能、免疫力和抗病力,且在添加量为1.5%时,效果最显著。     

两种乳酸菌对尼罗罗非鱼生长及免疫性能的影响

为研究饲料中添加植物乳杆菌(Lactobacillus plantarum)LP-37与戊糖片球菌(Pediococcus pentosaceus) PP-23对尼罗罗非鱼(Oreochromis niloticus)生长及免疫性能的影响,试验共分为7组:对照组C0,投喂基础饲料;L1-L3组,在基础饲料中添加不同浓度(1.0×10⁶ CFU/g、1.0×10⁷ CFU/g、1.0×10⁸ CFU/g)植物乳杆菌LP-37;P1-P3组,在基础饲料中添加不同浓度(1.0×10⁶ CFU/g、1.0×10⁷ CFU/g、1.0×10⁸ CFU/g)戊糖片球菌PP-23。结果表明:除L1组外,其余试验组末均重(123.22～158.60) g较C0组(118.16±4.88) g均显著增高(P<0.05);除L1组与P1组外,其余试验组增重率(128.69%～156.20%)较C0组(108.16±0.45)%均显著提高(P<0.05),L3与P2...     

河豚卵处理物对吉富罗非鱼生长性能、血液生化及抗菌感染的影响

为了探究不同添加水平的河豚(Takifugu rubripes)卵处理物对吉富罗非鱼(Oreochromis niloticus)生长性能、血液生化以及海豚链球菌感染后的存活率的影响,试验选取525尾平均体重为(0.66±0.01)g的吉富罗非鱼,随机分为7组,每组3个重复,每重复25尾鱼,依次饲喂河豚卵处理物添加水平为0、0.2%、0.4%、0.8%、1.6%、3.2%和6.4%的饲料,养殖8周。结果显示:随着河豚卵处理物添加水平的提高,试验鱼体增重、特定生长率、肝体比和脏体比均先上升后下降最后趋于稳定,饲料转化率显著降低,各组肝体比无显著差异,0.8%河豚卵处理物添加组罗非鱼的脏体比显著高于其他组。随着河豚卵处理物添加水平的提高,罗非鱼血液中红细胞、血红蛋白、红细胞比容含量显著下降,白细胞数量呈逐渐上升的趋势;罗非鱼血清中总胆固醇和血糖含量显著下降,血清中溶菌酶含量呈逐渐下降的趋势。罗非鱼腹腔注射5×10~7 CFU/mL海豚链球菌后,各组罗非鱼24 h内均未出现死亡;感染48～96 h,对照组罗非鱼累计存活率最低;感染192 h后,6.4%河豚卵处理物添加组罗非鱼累计存活率显著高于对照组。结果表明:饲料中添加河豚卵处理物对吉富罗非鱼的生长性能、血液生化指标产生影响,增强了罗非鱼抗海豚链球菌病感染的能力。     

饲料中添加乳酸菌对方斑东风螺稚螺生长、肌肉组成与免疫功能的影响

为探讨饲料中添加乳酸菌对方斑东风螺稚螺生长、肌肉组成与免疫的影响,在基础饲料中添加三个不同浓度(1.2×10~7 CFU/100g、1.2×10~8 CFU/100g和1.2×10~9 CFU/100g)的乳酸菌,养殖初始体重为0.62g的方斑东风螺10周。结果显示,与对照组相比,饲料中添加乳酸菌对方斑东风螺稚螺的生长性能和肌肉组分均没有显著影响。饲料中添加1.2×10~7 CFU/100g乳酸菌能够显著提高方斑东风螺的超氧化物歧化酶(SOD)活性,但在添加量为1.2×10~8 CFU/100g和1.2×10~9 CFU/100g时SOD活性反而受到抑制;添加1.2×10~9 CFU/100g乳酸菌抑制酸性磷酸酶(ACP)活性;饲料中添加乳酸菌对过氧化氢酶(CAT)和碱性磷酸酶(AKP)活性没有显著影响。结果表明饲料中添加乳酸菌不影响方斑东风螺稚螺的生长性能和肌肉成分,添加量为1.2×10~7 CFU/100g时可一定程度上提高抗氧化活力,但较高的添加量对免疫功能会产生抑制作用。     

3种饲料添加剂对吉富罗非鱼幼鱼生长性能、消化酶活性和肝、肠组织结构的影响

为探讨中草药提取物、胆汁酸、微生态制剂对吉富罗非鱼(Oreochromis niloticus)生长性能、消化酶活性和肝、肠组织结构的影响,以基础饲料为对照,分别在每千克基础饲料中添加中草药提取物(1、2 g)、胆汁酸(0.1、0.2 g)、微生态制剂(1、2 g)配制成6种试验饲料,选取平均体质量为(32.75±5.75)g的吉富罗非鱼幼鱼483尾,随机分为7处理,每处理3平行,每平行23尾,共进行养殖实验45 d。结果显示:与对照组相比,低剂量使用此3种添加剂均有改善实验鱼生长和饲料利用的趋势。饲料中添加胆汁酸显著提高了吉富罗非鱼的增重率和特定增长率;饲料中添加微生态制剂组的吉富罗非鱼的肥满度最大,但各处理组之间差异不显著;中草药组的增重率与特定生长率与对照组之间无差异。饲料中添加微生态制剂和0.1 g/kg胆汁酸的实验鱼肠道脂肪酶活性显著高于中草药提取物2 g/kg的处理组;各组间的肠道淀粉酶、蛋白酶活性差异不显著。与对照组相比,各处理组实验鱼的前肠绒毛个数和肌肉层厚度有增加的趋势;添加1 g/kg微生态制剂的实验鱼中肠绒毛长度显著高于中草药组。结果表明:饲料中添加1 g/kg中草药提取物,0.2 g/kg胆汁酸,1 g/kg微生态制剂可在一定程度上提高吉富罗非鱼幼鱼的生长性能。     

5种乳酸菌对奥尼罗非鱼免疫和抗病力的影响

选用体质量约3．49 g的奥尼罗非鱼,分别投喂基础饲料（空白组）和添加相同水平的嗜酸乳杆菌、植物乳杆菌、戊糖乳杆菌、鼠李糖乳杆菌、粪肠球菌的试验饲料。饲料中的乳酸菌数分别为1．30×103、5．10×108、2．97×108、3．03×108、2．30×108、3．47×108 cfu／g ,养殖期为8周,试验结束后采集血液并留部分鱼进行72 h的嗜水气单胞菌感染试验,计算72 h内的存活率。结果显示,植物乳杆菌组、戊糖乳杆菌组、鼠李糖乳杆菌和粪肠球菌组血清碱性磷酸酶活性均有不同程度升高,其中戊糖乳杆菌组最高,且显著高于嗜酸乳杆菌组;鼠李糖乳杆菌和粪肠球菌组血清抗超氧阴离子自由基能力显著高于其他各组;鼠李糖乳杆菌组血清溶菌酶活性显著高于空白组。注射嗜水气单胞菌12 h后,各添加组罗非鱼存活率均高于空白组,24～48 h ,鼠李糖乳杆菌组和粪肠球菌组存活率显著高于空白组,72 h时鼠李糖乳杆菌组存活率显著高于空白组。饲料中添加乳酸菌能提高奥尼罗非鱼免疫、抗氧化能力及抗嗜水气单胞菌感染能力,其中鼠李糖乳杆菌和粪肠球菌效果显著。     

中草药复合微生态制剂对吉富罗非鱼(Oreochromis niloticus)生长、肠道菌群及抗病力的影响

通过在饲料中分别添加2×10~7 CFU/g的芽孢杆菌制剂、中草药芽孢杆菌制剂、复合微生态制剂和中草药复合微生态制剂,研究4种微生态制剂对吉富罗非鱼(Oreochromis niloticus)生长、肠道菌群及抗病力等的影响。结果显示:(1)饲料中添加4种微生态制剂均可显著提高罗非鱼的增重率(P0.05),对成活率和饲料利用率也有一定程度的提高(P0.05),而中草药复合微生态制剂对罗非鱼促进生长效果最佳。(2)饲料中添加4种微生态制剂可以显著提高罗非鱼肠道中的细菌总数、芽孢杆菌、乳酸杆菌和双歧杆菌数量(P0.05),大肠杆菌数量显著低于对照组(P0.05),说明饲料中添加一定量的4种微生态制均可改善罗非鱼的肠道菌群结构,以中草药复合微生态制剂的改善效果最佳。(3)经人工感染无乳链球菌后,罗非鱼对照组全部死亡,4个实验组只有部分死亡。鉴定发现,吉富罗非鱼的死亡均由感染无乳链球菌所致,试验组罗非鱼的免疫保护率分别为51.42%(B组)、58.62%(C组)、58.62%(D组)和68.93%(E组),以中草药复合微生态制剂组的免疫保护率最高。综上所述,在饲料中添加一定比例的中草药复合微生态制剂可以提高吉富罗非鱼生长指标、改善其肠道菌群结构和增加抗病力。     

饲料中添加谷胱甘肽对吉富罗非鱼生长、组织生化指标和非特异性免疫相关酶的影响

为研究饲料中添加谷胱甘肽(glutathione,GSH)对吉富罗非鱼(GIFT)生长性能、组织生化指标和非特异性免疫相关酶活性的影响,实验选用720尾体质量为(3.27±0.04)g的罗非鱼,随机分为6组,分别投喂基础饲料(对照组)和5种添加80、160、240、320和400 mg/kg GSH的试验饲料,养殖期为7周。结果显示,与对照组相比,320 mg/kg组罗非鱼的增重率(WGR)、特定生长率(SGR)、蛋白质沉积率(PDR)和肝脏RNA/DNA比值分别显著升高,饲料系数显著降低;各添加组罗非鱼肝体比高于对照组,但差异不显著。160～320mg/kg组罗非鱼粗蛋白和240～300 mg/kg组粗脂肪含量显著高于对照组,均在240 mg/kg组达到最高值。320 mg/kg组血清尿素氮(UN)含量与对照组相比显著降低。160～400mg/kg组血清和肝脏类胰岛素生长因子Ⅰ(IGF-Ⅰ)显著高于对照组和80 mg/kg组。240～400 mg/kg组血清溶菌酶(LZM)、320～400 mg/kg组肝脏LZM活性分别显著高于其它各组;与对照组相比,320 mg/kg组血清一氧化氮合成酶(NOS)活性显著升高;各添加组血清碱性磷酸酶(AKP)和酚氧化酶(PO)、肝脏AKP、酸性磷酸酶(ACP)和NOS活性均高于对照组,但差异不显著。结果表明,饲料中添加一定量的谷胱甘肽能显著提高吉富罗非鱼幼鱼的生长性能,提高全鱼粗蛋白与粗脂肪含量、血清和肝脏IGF-Ⅰ水平以及非特异性免疫相关酶活性。以增重率为评价指标,计算出吉富罗非鱼幼鱼饲料中谷胱甘肽的最适添加量为355.13 mg/kg。     

饲料中添加水飞蓟素对吉富罗非鱼生长性能、肝脏脂肪代谢酶和抗氧化能力的影响

为评估饲料中添加水飞蓟素对吉富罗非鱼生长性能、肝脏脂肪代谢酶和抗氧化能力的影响,分别在5组等氮等能的吉富罗非鱼饲料中添加不同水平的水飞蓟素[0(对照组)、100、200、400和800 mg/kg],饲喂初始体质量为(8.17±0.31)g的吉富罗非鱼幼鱼9周,测定实验鱼的生长性能、血清生化指标、肝脏脂肪代谢酶活性及抗氧化酶活性。结果显示,随饲料中水飞蓟素添加水平增加,吉富罗非鱼终末体质量、增重率、特定生长率和肥满度均呈先增加后降低的趋势,各指标均在100mg/kg实验组达到最大,100mg/kg实验组的饲料系数显著低于对照组。饲料中添加水飞蓟素显著降低了实验鱼肝体比和脏体比。100mg/kg实验组全鱼、肌肉和肝脏的粗脂肪含量显著低于对照组。血清谷草转氨酶和谷丙转氨酶活性随水飞蓟素水平增加呈先降低后升高的趋势,分别在200和100 mg/kg实验组达到最低,两组之间无显著性差异。饲料中添加水飞蓟素后,血清中的甘油三酯含量显著低于对照组;肝脏中的脂蛋白脂酶和总酯酶活性显著高于对照组;肝脏脂肪酶活性呈先上升后降低趋势,在200 mg/kg实验组最高,且与对照组有显著性差异。饲料中添加水飞蓟素后,肝脏谷胱甘肽过氧化物酶活性显著高于对照组,肝脏过氧化氢酶活性显著低于对照组;肝脏丙二醛含量显著低于对照组。100 mg/kg实验组罗非鱼的肝细胞界限清晰,无核偏移,空泡较少。研究表明,饲料中添加适量水飞蓟素提高了吉富罗非鱼生长性能,调节了肝脏脂肪代谢酶活性和抗氧化能力,对防治肝脏细胞损伤有积极作用。在本实验条件下,推荐吉富罗非鱼幼鱼饲料中水飞蓟素添加水平为100 mg/kg。     

中草药复合微生态制剂对吉富罗非鱼(Oreochromis niloticus)生长、肠道菌群及抗病力的影响

通过在饲料中分别添加2×107 CFU/g的芽孢杆菌制剂、中草药芽孢杆菌制剂、复合微生态制剂和中草药复合微生态制剂,研究4种微生态制剂对吉富罗非鱼(Oreochromis niloticus)生长、肠道菌群及抗病力等的影响。结果显示：(1)饲料中添加4种微生态制剂均可显著提高罗非鱼的增重率(P<0.05),对成活率和饲料利用率也有一定程度的提高(P>0.05),而中草药复合微生态制剂对罗非鱼促进生长效果最佳。(2)饲料中添加4种微生态制剂可以显著提高罗非鱼肠道中的细菌总数、芽孢杆菌、乳酸杆菌和双歧杆菌数量(P<0.05),大肠杆菌数量显著低于对照组(P<0.05),说明饲料中添加一定量的4种微生态制均可改善罗非鱼的肠道菌群结构,以中草药复合微生态制剂的改善效果最佳。(3)经人工感染无乳链球菌后,罗非鱼对照组全部死亡,4个实验组只有部分死亡。鉴定发现,吉富罗非鱼的死亡均由感染无乳链球菌所致,试验组罗非鱼的免疫保护率分别为51.42%(B组)、58.62%(C组)、58.62%(D组)和68.93%(E组),以中草药复合微生态制剂组的免疫保护率最高。综上所述,在饲料中添加一定比例的中草药复合微生态制剂可以提高吉富罗非鱼生长指标、改善其肠道菌群结构和增加抗病力。     

BALB/c小鼠用于评价尼罗罗非鱼无乳链球菌毒力的研究

实验采用BALB/c小鼠作为实验动物,旨在建立尼罗罗非鱼无乳链球菌毒力测定的BALB/c小鼠模型。BALB/c小鼠经腹腔注射尼罗罗非鱼源无乳链球菌建立感染模型,比较了尼罗罗非鱼源无乳链球菌分别感染尼罗罗非鱼和小鼠的LD_(50)差异,分别测定了不同毒力尼罗罗非鱼无乳链球菌对尼罗罗非鱼和小鼠的毒力。结果显示,小鼠经腹腔注射无乳链球菌,在24 h内出现死亡现象,且对小鼠脑、肝脏、脾脏、肾脏等组织造成损伤。3次测定尼罗罗非鱼无乳链球菌TFJ0901对尼罗罗非鱼和小鼠LD_(50)分别为7.7×10~7、2.2×10~8、3.5×10~9 CFU/mL和405、361、419 CFU/只。将无乳链球菌TFJ0901和THN0901感染尼罗罗非鱼(1.0×10~7 CFU/mL)和小鼠(100 CFU/只),尼罗罗非鱼和小鼠存活率分别为100%、6.7%±5.8%和100%、0,其存活率都具有显著性差异。将无乳链球菌TFJ0901和TFJ-F感染尼罗罗非鱼(3.0×10~8 CFU/mL)和小鼠(2 500 CFU/只),尼罗罗非鱼的存活率分别为73.3%±11.5%和80.0%±10.0%,存活率差异不显著,小鼠存活率分别为13.3%±11.5%和100.0%,存活率具有显著性差异。研究表明,本实验成功建立了BALB/c小鼠作为尼罗罗非鱼源无乳链球菌毒力测定的稳定模型,测定不同毒力的尼罗罗非鱼源无乳链球菌对小鼠毒力与对尼罗罗非鱼毒力一致,且该模型能够区分尼罗罗非鱼模型难以区分的毒力相近的无乳链球菌。     

灿烂弧菌对厚壳贻贝免疫指标和消化酶活性的影响

为探究灿烂弧菌对厚壳贻贝免疫指标和消化酶活性的影响,用1×10~6、1×10~7、1×10~8个/mL 3个浓度的灿烂弧菌刺激厚壳贻贝,探讨弧菌刺激后厚壳贻贝的一氧化氮合酶(NOS)、一氧化氮(NO)、超氧化物歧化酶(SOD)、丙二醛(MAD)等免疫指标和淀粉酶、蛋白酶等消化酶的变化情况。结果显示,灿烂弧菌刺激48 h后,厚壳贻贝足、鳃、消化腺等组织中仅消化腺的NOS活性较对照组有显著升高,且酶活性随初始细菌浓度的升高而升高,故选用消化腺来测定灿烂弧菌刺激后72 h内的免疫指标和消化酶活性的变化。灿烂弧菌刺激后,48 h内NOS活性较对照组均显著升高。NO含量较对照组均显著升高,与NOS显现相同变化趋势。SOD活性在各浓度灿烂弧菌刺激下较对照组均显著升高,而MAD含量在实验组中含量显著低于对照组。淀粉酶活性在实验组中显著低于对照组,总体呈现先下降后升高的趋势。蛋白酶活性在各实验组中均呈现先升高后下降的趋势。研究表明,灿烂弧菌对厚壳贻贝免疫指标和蛋白酶活性的升高有诱导作用,但对蛋白酶的活性有抑制作用。本研究初步探明了厚壳贻贝对灿烂弧菌的免疫应答机制,为进一步研究灿烂弧菌和厚壳贻贝相互作用机制以及厚壳贻贝免疫机制奠定了基础。     

尼罗罗非鱼无乳链球菌Sip蛋白乳酸菌活载体口服疫苗的研制及其免疫效果

链球菌病是威胁我国罗非鱼养殖产业健康发展的重要病害之一。为研制出免疫效果好、操作简便的罗非鱼链球菌病疫苗,本研究构建重组表达无乳链球菌Sip蛋白的穿梭质粒pNZ8124-Sip,通过酶切和测序验证后电转化乳酸乳球菌NZ9000,获得能够诱导重组表达无乳链球菌Sip蛋白的乳酸菌活菌载体疫苗。采用SPS-PAGE电泳摸索最佳诱导浓度和诱导时间以获得最大表达量,通过镍柱纯化目的蛋白并进行Western blot检测;利用不同浓度的重组乳酸菌活载体疫苗灌胃口服免疫尼罗罗非鱼,采用间接ELISA法测定免疫后血清抗体水平变化,通过人工腹腔注射感染无乳链球菌获得相对免疫保护率。研究结果显示,构建的重组乳酸乳球菌可通过nisin诱导表达大小为48 ku特异性蛋白,与目的蛋白大小一致;PAGE电泳显示,重组蛋白主要以可溶蛋白和包涵体2种形式存在,其中胞内可溶性蛋白浓度达7.65 mg/mL;诱导表达的最佳条件为100 ng/mL nisin诱导6 h;Western blot检测结果显示,诱导蛋白可与鼠抗His标签抗体特异性结合。口服免疫结果显示,中浓度组(2.24×10~(10) CFU/mL)和低浓度组(2.24×10~9 CFU/mL)免疫2次能够显著提高尼罗罗非鱼的血清抗体水平和抗无乳链球菌感染能力,中浓度免疫组的相对免疫保护率最高为41.0%。本研究可为罗非鱼链球菌病口服疫苗的研究奠定基础,具有广阔的应用前景。     

Delivery of HUFA, probionts and biomedicine through bioencapsulated Artemia as a means to enhance the growth and survival and reduce the pathogenesity in shrimp Penaeus monodon postlarvae

One of the major problems in the shrimp culture industry is the difficulty in producing high-quality shrimp larvae. In larviculture, quality feeds containing a high content of highly unsaturated fatty acids (HUFA) and ingredients that stimulate stress and disease resistance are essential to produce healthy shrimp larvae. In the present study, Penaeus monodon postlarvae (PL15) were fed for 25 days on an unenriched Artemia diet (control; A) or on a diet of Artemia enriched with either HUFA-rich liver oil of the trash fish Odonus niger (B), probionts [Lactobacillus acidophilus (C1) or yeast-Saccharomyces cerevisiae (C2)] or biomedicinal herbal products (D) that have anti-stress, growth-promoting and anti-microbial characteristics. P. monodon postlarvae fed unenriched Artemia exhibited the lowest weight gain (227.9 ± 8.30 mg) and specific growth rate (9.95 ± 0.05%), while those fed the HUFA-enriched Artemia (B) exhibited the highest weight gain and specific growth rate (362.34 ± 12.56 mg and 11.77 ± 0.08%, respectively). At the end of the 25-day rearing experiment, the shrimp postlarvae (PL40) were subjected to a salinity stress study. At both low and high (0 and 50‰) salinities, the group fed the control diet (A) experienced the highest cumulative mortality indices (CMI) 935.7 ± 2.1 and 1270.7 ± 3.1, respectively. Those fed diet D showed the lowest stress-induced mortality, and CMI were reduced by 31.1 and 32.3% under conditions of low and high salinity stress, respectively. A 10-day disease challenge test was conducted with the P. monodon postlarvae (PL40–PL50) by inoculating the shrimp with the pathogen Vibrio harveyi at the rate of 10⁵–10⁷ CFU/ml in all rearing tanks. P. monodon postlarvae fed probiont-encapsulated Artemia diets (C1 and C2) exhibited the highest survival (94.3 and 82.3%, respectively) and lowest pathogen load (V. harveyi) in hepatopancreas (5.2 × 10² ± 9.0 × 10 and 4.6 × 10² ± 9.0 × 10 CFU g⁻¹, respectively) and muscle (2.0 × 10² ± 6 × 10 and 1.7 × 10² ± 8.6 × 10 CFU g⁻¹, respectively) tissues. The shrimp that were fed the unenriched Artemia (Control; A) showed the lowest survival (26.33%) and highest bacterial load in the hepatopancreas (1.0 × 10⁵ ± 5 × 10³ CFU g⁻¹) and muscle (3.6 × 10⁴ ± 6 × 10² CFU g⁻¹). The shrimp fed the herbal product (D)-enriched Artemia also exhibited enhanced survival and reduced V. harveyi load in the tissues tested compared to the control diet (A) group. The results are discussed in terms of developing a quality larval feed to produce healthy shrimp larvae.     

Effect of a probiotic bacterium <Emphasis Type="Italic">Bacillus circulans</Emphasis> PB7 in the formulated diets: on growth,nutritional quality and immunity of <Emphasis Type="Italic">Catla catla</Emphasis> (Ham.)

Bacillus circulans PB7, isolated from the intestine of Catla catla, was evaluated for use as a probiotic supplement in the feeds for the fingerlings of Catla catla. The effect of supplement on the growth performance, feed utilization efficiency, and immune response was evaluated. Catla fingerlings (ave. wt. 6.48 ± 0.43 g) were fed diets supplemented with 2 × 10⁴ (feed C1), 2 × 10⁵ (feed C2), and 2 × 10⁶ (feed C3) B. circulans PB 7 cells per 100 g feed for 60 days at 5% of the body weight per day in two equal instalments in triplicate treatments. The control feed (CC) was not supplemented with the B. circulans. All the feeds were isocaloric and isonitrogenous. Fish fed with feed C2 displayed better growth, significantly (P ≤ 0.05) highest RNA/DNA ratio, a lower feed conversion ratio (FCR), and a higher protein efficiency ratio (PER) than the other experimental diets. Highest carcass protein and lipid was also observed in the fish fed C2 feed compared to the others. Significantly (P ≤ 0.05), highest protease was recorded in fish fed feed C2 (47.9 ± 0.016) and lowest in fish fed feed C3 (32.10 ± 0.009), where α-amylase activity did not differ significantly (P ≤ 0.05) beyond the lowest inclusion level. ALP, ACP, GOT, and GPT in the liver of Catla catla were the highest (P ≤ 0.05) in fish fed C2 feed. The highest TSP, albumin, and globulin was observed in fish treated with C2 feed after 60 days feeding trial, but the lowest glucose level was observed in the same treatment. After the feeding trial, the non-specific immunity levels and disease resistance of fish were also studied. Phagocytic ratio, phagocytic index, and leucocrit value were the highest in fish fed feed C2. After the feeding trial, the fish were challenged for 10 days by bath exposure to Aeromonas hydrophila (AH1) (10⁵c.f.u. ml⁻¹ for 1 h, and, after 7 days, 10⁷c.f.u. ml⁻¹ for 1 h). Highest survival percentage was observed in fish fed with feed C2 compared with only 6.66% in the controls, which indicated the effectiveness of B. circulans PB 7 in reducing disease caused by A. hydrophila.     

患腹水病牙鲆病原菌分离、鉴定及病原菌的特性

为确定引起河北北戴河地区养殖牙鲆患腹水病死亡的病原,本实验从患腹水病牙鲆体内分离到3株优势菌,对分离株进行生理生化鉴定和16S rRNA序列比对来确定分离株的生物学地位,进一步通过毒力基因(toxR、vhhA、vhhB)鉴定及组织病理学分析其病理特征,并通过人工回感实验分析分离株毒性。结果显示,通过生理生化实验及16S rRNA序列比对,确定此次从患病牙鲆中分离到的3株菌株均为哈维氏弧菌;经鉴定,3株分离株毒力基因(toxR、vhhA、vhhB)结果均为阳性,病理组织切片结果显示,该分离株对牙鲆多器官(肠、肾脏、脾脏和肝脏)均可造成不同程度的病理损伤,呈全身性感染;人工回感实验显示,菌株BDHYPFS-Y1G对牙鲆的半致死浓度为LD50=5.88×106 CFU/mL,低于自然状态毒性;药敏实验表明,3株分离株均对呋喃妥因高度敏感。本实验确认了此次牙鲆腹水病的病原菌,并初步研究了病原菌致病性以及药物敏感性,以期为牙鲆工厂化养殖疫病防控提供科学依据。     

凡纳滨对虾白便综合征发生与环境因子、机体免疫酶活性和微生物的相关性

为系统解析凡纳滨对虾白便综合征 (white feces syndrome，WFS)的发生与环境因子、微生物因子、宿主免疫力和水体微生物群落组成的关系。实验利用水体理化因子、可培养细菌、对虾机体免疫酶活性以及高通量测序等检测技术对健康与患WFS的池塘养殖凡纳滨对虾进行比较分析。结果显示，与健康组相比，患病池塘的水温、溶解氧 (DO)、pH、盐度等水质理化因子波动趋势相似，波动范围分别为26.1~29.0 °C、4.26~6.08 mg/L、8.39~8.73和40~49，患病组DO和盐度比健康组高；健康组对虾肝胰腺内可培养细菌和弧菌含量为1.19×10⁵~7.70×10⁵和8.8×10³~1.96×10⁴ CFU/g，弧菌占比为2%~16%，患病组对虾肝胰腺内可培养细菌和弧菌含量在3.80×10⁵~2.51×10⁶和2.02×10⁵~1.49×10⁶ CFU/g范围内，比健康组高15~113倍，弧菌占比在55%~70%。碱性磷酸酶 (AKP)、酸性磷酸酶 (ACP)、溶菌酶 (LZM)、超氧化物歧化酶 (SOD)和酚氧化酶 (PO)活性在健康组内为1.21~5.64、9.17~15.25、3.56~7.43、4.83~6.70及3.10~4.55 U/mg，在患病组内为2.12~5.39、19.22~26.96、19.73~26.85、3.00~4.14及7.76~9.21 U/mg。比较分析表明，WFS的发生与可培养细菌含量、弧菌占比、ACP、LZM、PO的相关性较强。高通量测序分析表明，患病组水体菌群结构的Ace和Chao指数呈一定程度下降趋势，PCoA指数偏离度较高，放线菌门、变形菌门相对丰度降低，拟杆菌门、蓝藻门相对丰度显著升高；RDA关联分析表明，盐度、溶解氧、虾体细菌、虾体弧菌、水体细菌是影响患病对虾水体菌群结构组成的重要因子。相关研究结果为解析养殖生产中对虾WFS发生机制提供数据支撑，并为WFS的临床防控奠定理论基础。     

Enrichment of Artemia nauplii with the probiotic yeast Saccharomyces boulardii and its resistance against a pathogenic Vibrio

Enrichment of Artemia nauplii with a known probiotic yeast Saccharomyces boulardii (SB) and its role in enhancing resistance against the pathogen Vibrio harveyi was investigated. SB was cultured, then fed to instar II Artemia nauplii in three different treatments; 10² (T1), 10³ (T2) and 10⁴ (T3) colony forming units (CFU) per ml in triplicate. The algae Nanochloropsis sp. was used as control diet. Survival and total count of CFU nauplii⁻¹ was observed on different media (Sabouraud, for enumerating yeasts, thiosulphate citrate bile salts sucrose, for enumerating Vibrio and seawater agar, for enumerating total aerobic flora) for each replication. Enhanced survival of nauplii was observed in treatments as compared to control. Results indicated that enrichment of SB in Artemia nauplii proceeded in a linear fashion, and up to 3500 CFU of SB could be detected in one nauplii at 10⁴ CFU ml⁻¹ treatment. No conclusive trend could be observed in the count of Vibrio and total aerobic flora due to treatment. Enriched nauplii were then challenged with the pathogen V. harveyi for 24 and 48 h at a concentration of 6.1 × 10⁶ CFU ml⁻¹. The survival counts at 48 h showed that the resistance of the nauplii was significantly (P < 0.01) improved in those fed with 10⁴ CFU  ml⁻¹ SB (90% survival rate after 48 h of challenge versus less than 40% for the infected control group without SB and treatments T1 and T2). This study shows that SB, which has been used for the first time in an aquatic live feed organism, has a profound beneficial effect on the nauplii by increasing its resistance to a pathogenic Vibrio infection. This revised version was published online in August 2006 with corrections to the Cover Date.     

珠江三角洲地区大口黑鲈池塘养殖系统的能值评估

为评价珠江三角洲地区大口黑鲈池塘养殖系统的生态经济性能,实验以能值理论为基础,定量分析该系统的能流和物流特点,通过建立能值评价指标体系,综合评估该系统的环境影响及可持续性。结果显示,大口黑鲈养殖系统投入的资源分为可更新资源(太阳能、风能、雨水能、地球循环能和河水能)和购买的外部资源(设施、苗种、电能、饵料、药品、劳动力、租金、维护费)两部分。养殖系统投入的总能值为4.51×10~(17)sej/(hm~2·a),其中可更新资源能值总和为1.24×10~(16) sej/(hm~2·a),占总投入能值的2.75%。河水能在可更新资源中所占比例最大,为9.77×10~(15) sej/(hm~2·a),占总投入能值的2.17%。购买的外部资源能值总和为4.38×10~(17) sej/(hm~2·a),占总投入能值的97.25%。饵料投入在购买的外部资源中所占比例最大,其能值为3.49×10~(17) sej/(hm~2·a),占总投入能值的77.33%,其次是劳动力,能值为2.29×10~(16) sej/(hm~2·a),占总投入能值的5.08%。大口黑鲈池塘养殖系统太阳能值转换率TR为2.18×106 sej/J,产出能值交换率EERY为2.028,能值产出率EYR为1.028,环境负载率ELR为35.39,能值持续性指数ESI为0.029,可持续性发展的能值指数EISD为0.059。大口黑鲈池塘养殖系统经济效益较高,但过多依赖购买的外部资源,对环境压力较大,可持续性较差。减少饵料投喂量、提高饵料利用率(如选择优质配合饲料及添加剂、改进投喂策略等)以及开展综合养殖是提高珠江三角洲地区大口黑鲈养殖系统持续性、减小环境负载率的有效途径。     

The effects of dietary probiotic Bacilli (Bacillus subtilis and Bacillus licheniformis) on growth performance,feed efficiency,body composition and immune parameters of whiteleg shrimp (Litopenaeus vannamei) postlarvae

The present study investigates the effects of dietary commercial Bacilli probiotic (Bacillus subtilis and Bacillus licheniformis) on the growth performance, feed efficiency, body composition and immune parameters of Litopenaeus vannamei. The L. vannamei postlarvae were supplied and acclimated (in 500‐L tanks) to laboratory conditions for 14 days. The shrimps were fed with diets containing 1 × 10⁴ and 1 × 10⁸ CFU/g probiotic Bacilli for 60 days. At the end of the feeding trial, growth performance parameters, body composition, serum biochemical parameters and the hemocytes count were evaluated. Shrimps fed diets supplemented with 1 × 10⁴ and 1 × 10⁸ CFU/g probiotic Bacilli showed improved weight gain, total length, specific growth rate, FCR and survival compared with the control group. The body composition studies revealed higher dry matter, crude protein and ash in shrimps fed with 1 × 10⁴ and 1 × 10⁸ CFU/g probiotic Bacilli. Also, dietary administration of 1 × 10⁴ and 1 × 10⁸ CFU/g probiotic Bacilli decreased serum glucose and cortisol levels. However, significantly increased total protein, lysozyme and hemocyte cell count were noticed in shrimps fed 1 × 10⁴ and 1 × 10⁸ CFU/g probiotic Bacilli. In general, the findings of this study proved that oral administration of 1 × 10⁴ and 1 × 10⁸ CFU/g commercial probiotic Bacilli improved growth performance, feed utilization and immune parameters in whiteleg shrimp (Litopenaeus vannamei).