2株消化道优势菌对凡纳滨对虾免疫酶活性和抗白斑综合征病毒感染力的影响

以凡纳滨对虾为研究对象,在基础饲料中分别添加从健康对虾消化道中分离纯化的优势菌菌株——美人鱼发光杆菌PC463和坚强芽孢杆菌PC465(菌含量≥1011CFU/g)的活菌和破碎菌各1 g/kg,观察其对凡纳滨对虾血淋巴免疫酶活性和抗WSSV感染保护率的影响。经过20 d养殖实验后发现,与对照组相比,饲料中添加坚强芽孢杆菌活菌的免疫组和添加美人鱼发光杆菌灭活菌的免疫实验,其凡纳滨对虾血淋巴中酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性在不同程度上有所提高,并显著高于对照组(P<0.05)。WSSV感染后饲料中添加坚强芽孢杆菌活菌的免疫组存活率(53%±12%)和添加美人鱼发光杆菌灭活菌的免疫组存活率(49%±15%)显著高于对照组(P<0.05)。结果表明:饲料中添加坚强芽孢杆菌活菌和美人鱼发光杆菌灭活菌可以在一定程度上提高凡纳滨对虾免疫酶活性和抗WSSV感染能力,上述有防病作用的益生菌株以饲料添加剂的方式应用于对虾养殖生产,可望成为对虾白斑病生物防治的有效途径之一。     

几株肠道益生菌对凡纳滨对虾非特异免疫力和抗病力的影响

以基础饲料为对照组,在基础饲料中分别添加坚强芽孢杆菌活菌(Bacillus firmus)、坚强芽孢杆菌活菌(1.0×108 CFU/g)+美人鱼发光杆菌(Photobacterium damsela)灭活菌(1％)、坚强芽孢杆菌活菌(1.0×108 CFU/g)+溶藻弧菌(Vibrio alginolyticus)灭活菌(1％)配制3种免疫饲料.每组3个重复,对个体质量为(3.2±0.26)g的凡纳滨对虾(Litopenaeus vannamei)进行了为期30d的养殖实验.每5d取样,以血清中的酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、一氧化氮合酶(NOS)、超氧化物歧化酶(SOD)和溶菌酶(UL)活性为免疫指标,探讨了肠道益生菌及其灭活菌体作为免疫制剂对凡纳滨对虾非特异性免疫水平的影响;在投喂免疫饲料后的第16天,按0.9 g/10尾剂量,直接投喂感染白斑综合征病毒(wssV)对虾病料,计算各实验组每天的累计死亡率,分析肠道益生菌及其灭活菌体作为免疫制剂对凡纳滨对虾病毒感染能力的影响.结果显示:添加益生菌的实验组对虾血清中SOD、ACP、AKP和NOS活性明显高于对照组(P＜0.05),特别是显著提高了对虾抗WSSV感染的能力.其中坚强芽孢杆菌活菌(1.0× 108 CFU/g)和美人鱼发光杆菌灭活菌(1％)实验组的抗病毒感染能力最强,感染WSSV 14 d后累计死亡率为10.71％;而对照组为64.28％.结论认为,饲料中添加肠道益生菌及其灭活菌体能提高凡纳滨对虾非特异性免疫水平和抵抗疾病的能力,有望作为新型对虾免疫增强剂应用于对虾养殖业.     

美人鱼发光杆菌对凡纳滨对虾非特异性 免疫功能及抗病力的影响

在基础饲料中分别添加0.1%和1%美人鱼发光杆菌灭活菌、0.1%美人鱼发光杆菌活菌配制成3种免疫实验饲料,以基础饲料为空白对照组饲料,每组设3个平行样。对个体质量为(4.83±0.36)g的凡纳滨对虾进行为期20 d的饲养实验,分别在0、5、10、15和20d进行取样,以血清中的酚氧化酶(PO)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、超氧化物歧化酶(SOD)和溶菌酶(UL)活性为免疫指标,探讨了美人鱼发光杆菌作为免疫制剂对凡纳滨对虾非特异性免疫效应的影响;在投喂免疫饲料后的第22天,按0.004 2 kg/kg体重的剂量,直接投喂对虾白斑综合征病毒(WSSV)病料,并记录累积死亡率。结果表明,美人鱼发光杆菌免疫实验组对凡纳滨对虾血清中PO、ACP、AKP、UL和SOD活性影响明显高于对照组,并且在饲料中添加美人鱼发光杆菌后,明显提高了对虾抵御WSSV感染的能力。其中0.1%美人鱼发光杆菌活菌实验组的抗病毒感染能力最强,WSSV感染14d内累计死亡率为63.3%±5.8%;而对照组为96.7%±3.3%。研究表明,美人鱼发光杆菌添加在对虾饲料中能提高凡纳滨对虾非特异性免疫水平,增强抵抗疾病的能力,将其作为对虾免疫增强剂具有良好的应用前景。     

饲料中补充蜡样芽孢杆菌(Bacillus cereus)生物膜对凡纳滨对虾(Litopenaeus vannamei)生长、抗病力及其肠道微生物组成的影响

为了研究外源补充蜡样芽孢杆菌(Bacillus cereus)生物膜对凡纳滨对虾(Litopenaeus vannamei)生长、抗病力及对其肠道微生物组成的影响,以基础饲料为空白组,在基础饲料中添加蜡样芽孢杆菌游离态活菌作为游离态组(活菌含量为108 CFU/g);在基础饲料中添加蜡样芽孢杆菌活菌生物膜作为生物膜组(活菌含量为108 CFU/g);每组8个平行.在养殖大棚暂养7d后,对凡纳滨对虾进行40 d的养殖实验.在第17天进行白斑综合征(White Spot Syndrome Virus,WSSV)攻毒实验;第22天进行副溶血弧菌(Vibrio parahaemolyticus)攻毒实验.期间在第1、5、10、15天采样,称重并测量体长计算生长速率.在第1、5、10、15、20、25天采样,并取其肠道内容物提取DNA,用16S rDNA序列V3+V4区高通量测序方法检测对虾肠道内微生物群落的结构及变化情况.结果显示,生物膜组和游离态组对虾体重、体长增长速率高于空白组,生物膜组与游离态组对虾体重、体长差异性不显著,生物膜组和游离态组对虾体重、体长与空白组相比差异性显著.实验中凡纳滨对虾肠道微生物由变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)、放线菌门(Actinobacteria)、厚壁菌门(Firmicutes)等组成,其中,变形菌门平均占到总量的94.0％.变形菌门中主要以弧菌属(Vibrio)、发光杆菌属(Photobacterium)、Octadecabacter等为主,生物膜组、游离态组、空白对照组弧菌属平均含量分别为34.65％、39.27％、58.00％.在WSSV攻毒实验中,生物膜组、游离态组、空白对照组平均累积死亡率分别为80.0％、77.0％、92.0％,各组差异性不显著(p＞0.05).在副溶血弧菌攻毒实验中,生物膜组、游离态组、空白对照组平均累积死亡率分别为61.3％、75.0％、77.3％,生物膜组与游离态组和空白对照组相比差异性显著(P＜0.05).研究表明,饲料中添加蜡样芽孢杆菌生物膜和游离态蜡样芽孢杆菌投喂凡纳滨对虾后,可改变对虾肠道的微生物组成,提高凡纳滨对虾生长速度、增强抗病能力,而且添加蜡样芽孢杆菌生物膜效果最明显.     

四种糖作为碳源对凡纳滨对虾生长、肠道消化酶及肠道菌群的影响

为研究4种常见的糖源对凡纳滨对虾生长、肠道消化酶及肠道菌群的影响,选取初始体质量为(0.36±0.02) g的凡纳滨对虾为研究对象,从多种糖类中选择常见的4种糖(葡萄糖、果糖、淀粉、蔗糖)作为糖源配制饲料,实验共计4个处理,每个处理3个重复,每个重复40尾虾,养殖周期为56 d,每种糖源的添加量为20%。结果显示,(1)凡纳滨对虾的特定生长率和蛋白质效率均存在显著性差异,淀粉组显著高于其他3组,但各组间的肝体比和含肉率无显著差异。全虾体成分,蔗糖组的粗脂肪显著低于其他3组,葡萄糖组的灰分显著高于其他3组,粗蛋白与水分各组间无显著差异。(2)各组间对虾肠道的脂肪酶活性,淀粉组与蔗糖组显著高于葡萄糖组及果糖组,胃蛋白酶活性方面,淀粉组显著高于其他3组。(3)各组间凡纳滨对虾肠道的OTU、Ace指数及Chao指数无显著差异,Shannon指数淀粉组及蔗糖组显著高于葡萄糖组,Simpson指数淀粉组显著低于蔗糖组,而蔗糖组又显著低于葡萄糖组。门水平上,变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)与放线菌门(Actinobacteria)的相对丰度差异显著,但疣微菌门(Verrucomicrobia)相对丰度各组间无明显差异;在纲水平上,各处理组间的对虾肠道菌群的差异性较显著,淀粉组对虾肠道中黄杆菌纲(Flavobacteriia)、γ变形菌纲(Gammaproteobacteria)、放线菌纲(Actinobacteria)及噬纤维菌纲(Cytophagia)是明显高于蔗糖组与葡萄糖组。研究表明,凡纳滨对虾对淀粉和蔗糖的吸收利用优于葡萄糖和果糖,同时,淀粉和蔗糖对凡纳滨对虾肠道菌群多样性存在一定的影响,淀粉和蔗糖为凡纳滨对虾饲料中的适宜糖源。     

饲料中益生菌对凡纳滨对虾生长、肠道菌群及部分免疫指标的影响

以初体质量为(0.11±0.00)g的凡纳滨对虾(Litopenaeus vannamei)为研究对象,在室外水泥池进行10周饲喂实验,研究饲料中添加单一益生菌和复合益生菌对凡纳滨对虾生长、消化酶、肠道和粪便菌群及部分免疫指标的影响.以基础饲料为对照组,在基础饲料中分别添加地衣芽孢杆菌(Bacillus licheniformis)、枯草芽孢杆菌(Bacillus subtilis)、虾乐333(复合益生菌)、地衣芽孢杆菌+枯草芽孢杆菌(1:1)、虾乐333+地衣芽孢杆菌+枯草芽孢杆菌(1:1:1)各1 g/kg,配制5种实验饲料.实验结果显示,复合益生菌饲喂组对虾的特定生长率显著高于对照组(P<0.05),而单一益生菌饲喂组与对照组差异不显著(P<0.05).饲料中添加益生菌能使凡纳滨对虾肠道蛋白酶和淀粉酶活力升高,其中复合益生菌饲喂组淀粉酶活力显著高于对照组(P<0.05).与对照组比,饲料中添加益生菌显著降低了肠道及粪便弧菌数(P<0.05),不同程度提高了凡纳滨对虾血清蛋白浓度、酚氧化酶活力、溶菌酶活力和总抗氧化力,其中添加复合益生菌能使血清蛋白浓度及溶菌酶活力较对照组显著提高(P<0.05).上述结果表明,与饲喂单一益生菌相比,复合益生菌对凡纳滨对虾的生长、免疫力具有更好的促进作用.     

高通量测序法分析两株益生菌对凡纳滨对虾肠道菌群结构的影响

为分析荚膜红细菌和蜡样芽孢杆菌2株益生菌对凡纳滨对虾肠道菌群结构的影响,本实验对凡纳滨对虾进行为期30 d的养殖饲喂实验,饲喂后期利用高通量测序凡纳滨对虾肠道微生物的16S rDNA V4区,来分析不同益生菌饲喂后凡纳滨对虾肠道菌群的结构特征,并结合对虾体质量增加率和攻毒后累计死亡率的宏观指标来进行分析。结果显示：①空白组（CK）、荚膜红细菌组（Rc）和蜡样芽孢杆菌组（Bc）样品OTU范围为374~506,其中CK组对虾肠道菌群OTU数量最低,饲喂益生菌后的2组对虾肠道菌群中OTU数量相对较高;②在门分类水平上,3组的变形菌门数均为最多,CK组主要为变形菌门和少量拟杆菌门,Bc组主要有变形菌门、拟杆菌门、无壁细菌门、厚壁细菌门和假单胞菌,Rc组主要是变形菌门、拟杆菌门、厚壁菌门、酸杆菌门、放线细菌、梭杆菌门和蓝细菌;③稀释曲线和Shannon指数结果可见,CK组样品物种丰度和复杂度最低,且Bc组样品丰度和复杂度相对较高;④PCA分析发现,Rc组和CK组样品微生物组成较为接近,结合宏观对虾体质量增加率和攻毒后累计死亡率的结果分析,可见相较于荚膜红细菌,蜡样芽孢杆菌对凡纳滨对虾肠道微生物菌群影响更显著,且益生效果更佳。研究表明,饲喂益生菌可以扩增对虾肠道微生物菌群丰度,并能抑制弧菌属等有害菌群的生长,提高对虾体质量增加率并降低死亡率,从而达到益生效果,其中以饲喂蜡样芽孢杆菌菌株效果更佳。     

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

为了研究外源补充蜡样芽孢杆菌对凡纳滨对虾生长及对其肠道微生物组成的影响,以基础饲料为空白组,在基础饲料中添加蜡样芽孢杆菌活菌作为实验组(活菌含量为108 CFU/g),每组4个平行,在养殖场对凡纳滨对虾自幼苗至成虾进行一个完整养殖周期的养殖实验。选取养殖场中8个光照、位置等条件基本相同的养殖池分为2组,空白组投喂基础饲料,实验组投喂免疫饲料,养殖周期95 d。期间在第45、52、59、66、73天采样,称重并计算生长速率,并取其肠道内容物提取DNA,用16S r DNA序列V4区高通量测序方法检测对虾肠道内微生物群落的结构及变化情况。结果显示:(1)投喂免疫饲料的实验组对虾生长速度比空白组平均快15.2%。(2)健康的凡纳滨对虾肠道微生物以变形菌门和拟杆菌门含量高,厚壁菌门、疣微菌门、浮霉菌门、放线菌门含量较低。(3)空白组变形菌门平均含量(68.30%),明显高于实验组平均含量(57.94%);空白组拟杆菌门平均含量(23.58%)明显低于实验组平均含量(30.06%)。(4)变形菌门中以弧菌属、Anaerospora、小红卵菌属为主,空白组弧菌属平均占总含量的5.40%,实验组平均占总含量的1.94%;拟杆菌门中以异养硝化菌属、粘着杆菌属为主,实验组异养硝化菌属平均占总含量的8.12%,空白组平均占含量的5.56%。研究表明,饲料中添加蜡样芽胞杆菌投喂凡纳滨对虾后可改变对虾肠道的微生物组成,提高对虾生长速度。     

酵母培养物和芽孢杆菌对凡纳滨对虾生长、蛋白酶活性和免疫性能的影响

研究了饲料中添加酵母培养物(益康XP)和芽孢杆菌(Bacillus)制剂对凡纳滨对虾(Litopenaeusvannamei)生长、肌肉成分、蛋白酶活性和免疫功能的影响。在基础饲料(对照组)中分别添加0.075%益康XP、0.100%益康XP、0.200%芽孢杆菌,饲养均重(5.57±0.21)g的凡纳滨对虾45 d。结果表明,添加0.075%益康XP、0.100%益康XP、0.200%芽孢杆菌分别提高凡纳滨对虾增重率17.24%、12.29%、20.18%,降低饲料系数14.28%、9.77%、12.03%,提高蛋白质效率16.49%、10.05%、12.98%(P<0.05);添加酵母培养物和芽孢杆菌对凡纳滨对虾肌肉组成成分没有影响(P>0.05);0.075%益康XP组、0.100%益康XP组、0.200%芽孢杆菌组肝胰脏蛋白酶活性较对照组分别提高25.24%、15.22%、54.92%(P<0.05);各试验组肠蛋白酶活性与对照组相比有所提高,但均未达到显著水平(P>0.05);0.100%益康XP、0.200%芽孢杆菌组凡纳滨对虾血清酚氧化酶活性、溶菌酶活性和超氧化物歧化酶活性均显著高于对照组(P<0.05);攻毒实验表明,感染溶藻弧菌后,芽孢杆菌组对虾第1、3、4天的累积死亡率均显著低于对照组(P<0.05),第4天时的免疫保护率达到27.1%,而添加益康XP对累积死亡率无显著影响(P>0.05)。综上所述,饲料中添加酵母培养物益康XP和芽孢杆菌均能改善凡纳滨对虾生长性能和消化功能,芽孢杆菌能增强凡纳滨对虾对溶藻弧菌的抗感染功能。     

饲料中补充蜡样芽孢杆菌(Bacillus cereus)生物膜对凡纳滨对虾(Litopenaeus vannamei)生长、抗病力及其肠道微生物组成的影响

为了研究外源补充蜡样芽孢杆菌(Bacillus cereus)生物膜对凡纳滨对虾(Litopenaeus vannamei)生长、抗病力及对其肠道微生物组成的影响,以基础饲料为空白组,在基础饲料中添加蜡样芽孢杆菌游离态活菌作为游离态组(活菌含量为10~8 CFU/g);在基础饲料中添加蜡样芽孢杆菌活菌生物膜作为生物膜组(活菌含量为10~8 CFU/g);每组8个平行。在养殖大棚暂养7 d后,对凡纳滨对虾进行40 d的养殖实验。在第17天进行白斑综合征(White Spot Syndrome Virus,WSSV)攻毒实验;第22天进行副溶血弧菌(Vibrio parahaemolyticus)攻毒实验。期间在第1、5、10、15天采样,称重并测量体长计算生长速率。在第1、5、10、15、20、25天采样,并取其肠道内容物提取DNA,用16S r DNA序列V3+V4区高通量测序方法检测对虾肠道内微生物群落的结构及变化情况。结果显示,生物膜组和游离态组对虾体重、体长增长速率高于空白组,生物膜组与游离态组对虾体重、体长差异性不显著,生物膜组和游离态组对虾体重、体长与空白组相比差异性显著。实验中凡纳滨对虾肠道微生物由变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)、放线菌门(Actinobacteria)、厚壁菌门(Firmicutes)等组成,其中,变形菌门平均占到总量的94.0%。变形菌门中主要以弧菌属(Vibrio)、发光杆菌属(Photobacterium)、Octadecabacter等为主,生物膜组、游离态组、空白对照组弧菌属平均含量分别为34.65%、39.27%、58.00%。在WSSV攻毒实验中,生物膜组、游离态组、空白对照组平均累积死亡率分别为80.0%、77.0%、92.0%,各组差异性不显著(P0.05)。在副溶血弧菌攻毒实验中,生物膜组、游离态组、空白对照组平均累积死亡率分别为61.3%、75.0%、77.3%,生物膜组与游离态组和空白对照组相比差异性显著(P0.05)。研究表明,饲料中添加蜡样芽孢杆菌生物膜和游离态蜡样芽孢杆菌投喂凡纳滨对虾后,可改变对虾肠道的微生物组成,提高凡纳滨对虾生长速度、增强抗病能力,而且添加蜡样芽孢杆菌生物膜效果最明显。     

Effect of Three Probiotics Administered Through Live Feed on Digestive Enzyme Activity in California Halibut,Paralichthys californicus,Larvae

The objective of this study was to evaluate the effect of probiotics as digestive system maturation promoters in larvae of the California halibut, Paralichthys californicus, during ontogeny. Three commercial probiotics: Levucell^®, Bactocell^®, and live yeast, were supplied at a concentration of 50 mg/L, using enriched rotifers and Artemia nauplii as vectors. Total and specific activities of trypsin, leucine‐aminopeptidase (LAP), lipase, chymotrypsin, and pepsin were assessed throughout the development. Growth in terms of length was not significantly different between treatments. Total pepsin and chymotrypsin activity were significantly higher at the end of the experiment, 46‐d posthatch (dph) in larvae supplied with Bactocell. However, no significant effect of the probiotic was detected for trypsin, lipase, and LAP. On the basis of the results, California halibut larvae seemed to have adequate alkaline proteolytic activity early in development, whereas acid proteolytic activity was not detected until the complete formation of a functional stomach. The feeding protocol and probiotics evaluated were unable to improve the morphological development of digestive system or the digestive enzyme activity early on. Nevertheless, the increase in activity of pepsin and chymotrypsin observed at 46 dph suggests a potential use of these probiotics once the metamorphosis is completed, and thus warrants further evaluation.     

鱼类消化酶活性与其体长、体重和水质的相关性研究

对主养草鱼(Ctenopharyngodon idellus)池塘中4种鱼类消化器官的胃蛋白酶、胰蛋白酶、脂肪酶和淀粉酶的活性与鱼体长、体重及主要水质指标的相关性进行了研究。结果显示,草鱼肠道和肝胰脏的胃蛋白酶与水温具有显著正相关(P<0.05),而草鱼肠道胰蛋白酶、肠道和肝胰脏的脂肪酶以及鲢(Hypophthalmichthys molitrix)肝胰脏中淀粉酶与水温呈负相关(P>0.05)。4种鱼的消化酶活性与总氮和总磷呈正相关。匙吻鲟(Polyodon spathula)的脂肪酶和淀粉酶活性与化学耗氧量呈负相关;除鲢外,其他3种鱼的胃蛋白酶活性均与水体硬度和碱度呈正相关,而胰蛋白酶、脂肪酶和淀粉酶均与水体硬度和碱度呈负相关。草鱼和鳙(Aristichthys nobilis)肠道胰蛋白酶、脂肪酶和淀粉酶以及匙吻鲟消化道和肝脏胰蛋白酶与鱼的体长、体重均具有显著的正相关。     

Development of digestive enzyme activity in spotted rose snapper,Lutjanus guttatus (Steindachner, 1869) larvae

We describe digestive enzyme activity during the larval development of spotted rose snapper, Lutjanus guttatus. Trypsin, chymotrypsin, leucine aminopeptidase, pepsin, amylase, lipase, and acid and alkaline phosphatase activities were evaluated using spectrophotometric techniques from hatching through 30 days. The spotted rose snapper larvae present the same pattern of digestive enzyme activity previously reported for other species in which pancreatic (i.e., trypsin, chymotrypsin, amylase, and lipase) and intestinal (i.e., acid and alkaline phosphatases and leucine aminopeptidase) enzymatic activities are present from hatching allowing the larvae to digest and absorb nutrients in the yolk-sac and live prey by the time of first feeding. The digestive and absorption capacity of the spotted rose snapper increases during the larval development. A significant increase in individual activity of all enzymes occurs at 20 DAH, and around 25 DAH, the juvenile-type of digestion is observed with the appearance of pepsin secreted by the stomach, suggesting that maturation of the digestive function occurs around 20–25 DAH. Our results are in agreement with a previous suggestion that early weaning may be possible from 20 DAH. However, the patterns of enzymatic activities reported in our study should be considered during the formulation of an artificial diet for early weaning of the spotted rose snapper.     

The effects of seasonal,sex and size on the digestive enzyme activities in freshwater crayfish (Astacus leptodactylus Eschscholtz, 1823)

The effects of seasonal, sex and size on digestive enzyme activities of Astacus leptodactylus in natural habitat were investigated in the present study. The freshwater crayfish were sampled 584 individual as seasonally from Egirdir Lake. Ten male and 10 female individuals were sampled randomly from caught crayfish in each two different size ranges (4.5–7.0, 7.1–9.9 cm) and each season. The gastrointestinal tract, gastric, intestine and midgut gland were individually dissected on ice for enzyme analysis and stomach contents. At the end of the study, amylase activities were affected by interaction of season × sex × size in intestinal, season × sex in midgut gland. However, any interaction was not determined in amylase activities of the gastric. Lipase activities were affected by interaction of season × sex × size in intestinal and gastric, season × sex in midgut gland. The protease activities in each organ were affected by interaction of season, sex and size (p ? .05). In all organs, amylase and lipase activities were similar. However, the highest protease activity was in stomach while the lowest protease activity was the intestine. The protease activities were higher than amylase and lipase activities. There is a positive correlation between protease enzyme activities in gastric with gonad maturation and active feeding period. It can be said based on data of digestive system and high protease activity that A. leptodactylus is an omnivore species in need high protein. In addition, the reason for the low levels of lipase activity may be A. leptodactylus not prefer high fat foods or not fed high fat foods.     

Developmental changes in digestive enzyme activity in American shad, <Emphasis Type="Italic">Alosa sapidissima</Emphasis>, during early ontogeny

In order to assess the digestive physiological capacity of the American shad Alosa sapidissima and to establish feeding protocols that match larval nutritional requirements, we investigated the ontogenesis of digestive enzymes (trypsin, amylase, lipase, pepsin, alkaline phosphatase, and leucine aminopeptidase) in larvae, from hatching to 45 days after hatching (DAH). We found that all of the target enzymes were present at hatching, except pepsin, which indicated an initial ability to digest nutrients and precocious digestive system development. Trypsin rapidly increased to a maximum at 14 DAH. Amylase sharply increased until 10 DAH and exhibited a second increase at 33 DAH, which coincided with the introduction of microdiet at 30 DAH, thereby suggesting that the increase was associated with the microdiet carbohydrate content. Lipase increased until 14 DAH, decreased until 27 DAH, and then increased until 45 DAH. Pepsin was first detected at 27 DAH and then sharply increased until 45 DAH, which suggested the formation of a functional stomach. Both alkaline phosphatase and leucine aminopeptidase markedly increased until 18 DAH, which indicated intestinal maturation. According to our results, we conclude that American shad larvae possess the functional digestive system before mouth opening, and the significant increases in lipase, amylase, pepsin, and intestinal enzyme activities between 27 and 33 DAH suggest that larvae can be successfully weaned onto microdiets around this age.     

地衣芽孢杆菌对大泷六线鱼生长、肠道消化酶、血清非特异性免疫及抗病力的影响

本研究探讨饲料中添加地衣芽孢杆菌(Bacillus licheniformis)对大泷六线鱼(Hexagrammos otakii)生长、肠道消化酶、血清非特异性免疫及抗病力的影响。选取平均体重为(22.0±2.0) g的大泷六线鱼270尾,随机分成3组(对照组和2个不同菌剂水平的实验组),每组3个平行,每个平行 30尾鱼。对照组投喂仅有蛋白液包裹的基础饲料,实验组投喂含活菌量达5×107 CFU/g(0.5%)和1×108 CFU/g(1.0%)地衣芽孢杆菌的实验饲料。投喂50 d后进行致病菌杀鲑气单胞菌(Aeromonas salraonicida)攻毒感染,测定14 d内累积死亡率。结果显示,实验组饲料中地衣芽孢杆菌可显著提高大泷六线鱼的特定生长率(SGR) (P<0.05)。与对照组比较,0.5%和1.0%地衣芽孢杆菌均可提高大泷六线鱼血清中SOD、CAT及T-AOC活性(P<0.05),降低MDA含量(P<0.05);GSH-Px活性在1.0%实验组较对照组下降,而0.5%实验组较对照组提高。地衣芽孢杆菌实验组鱼血清GS、MDH和HK活性均得到提高(P<0.05),0.5%和1.0%不同剂量组的提高程度不同。地衣芽孢杆菌实验组鱼血清中AST和ALT活性较对照组均降低(P<0.05),CHE和ADA活性较对照组有提高(P<0.05)。实验组鱼肠道胰蛋白酶、淀粉酶和脂肪酶活性均有不同程度的提高,且1.0%实验组活性最强(P<0.05)。攻毒实验结果显示,1.0%地衣芽孢杆菌组大泷六线鱼14 d内累积死亡率仅有35.55% (P<0.05)。由此可见,饲料中添加地衣芽孢杆菌可促进大泷六线鱼生长,提高其肠道消化酶活性及非特异性免疫酶活性,增强其对杀鲑气单胞菌的抵抗能力。     

Cross‐effects of dietary probiotic supplementation and rearing temperature on growth performance,digestive enzyme activities,cumulative mortality and innate immune response in seabass (Dicentrarchus labrax)

Probiotic influence on fish immune response and digestive capacity is extensively discussed in aquaculture. In this experiment, a feeding trial was carried out for 100 days to evaluate the cross‐effects of probiotic supplementation and rearing temperature (17, 20 and 23°C) in juvenile seabass (Dicentrarchus labrax). The experimental diet was supplemented with a commercial probiotic blend (Biomin AquaStar Growout) at 3 g/kg diet (5.23 × 10⁸ CFU/kg diet), and tested against a non‐supplemented diet (control). Growth performance and innate immune responses were analysed at 70 and 100 days of feeding, whereas digestive enzyme activities were determined at 100 days of feeding. At the end of the feeding trial, fish were subjected to a handling stress and cumulative mortality was recorded. Digestive enzyme activities were influenced by temperature, with α‐amylase and lipase activities peaking at the higher temperature (23°C) and trypsin at the lower temperature (17°C). Immune parameters showed a significant temperature versus feeding duration effect, with complement system (ACH50) and peroxidase peaking at 70 and 100 days of feeding, respectively. Poststress cumulative mortality was higher at the lowest temperature (17°C), especially in fish fed the control diet. In conclusion, water temperature was the main variable affecting the studied parameters, whereas the dietary probiotic supplementation had influence on the chymotrypsin activity and survival rate in seabass reared at 17°C.     

Effect of dietary administration of Psychrobacter sp. on the growth,feed utilization,digestive enzymes and immune responses of grouper Epinephelus coioides

A 60‐day feeding trial was conducted to assess the effect of incorporating dietary Psychrobacter sp., one predominant bacterium with antagonistic activity in the gut of fast‐growing grouper Epinephelus coioides, on the growth, feed utilization, digestive enzymes and immune responses of E. coioides at regular monthly intervals. The fish were fed with control diet (without probiont) or probiotic diet supplemented with 1.0 × 10⁸ CFU g^?1Psychrobacter sp. No significant improvement in weight gain (WG) and specific growth ratio (SGR) were observed in the probiotic group, but a significant improvement in feed conversion ratio (FCR) was observed in fish fed the probiotic diet for 60 days. Although not statistically significant, enhanced hepatopancreatic protease and lipase activities and intestinal amylase activity were observed in probiotic group at day 60. A slightly increase in phagocytic activity and phagocytic index was observed in fish fed probiotic diet for 60 days. Serum lysozyme activity showed no significant differences between the probiotic group and the control group during the whole experiment period, while an increasing trend of superoxide dismutase (SOD) activity in the probiotic group was observed. Serum complement component 3 (C3) level of the probiotic group was higher than that of control in the whole feeding period, while complement component 4 (C4) level of the probiotic group was significantly higher than that of the control at day 60. The results of the current study firstly demonstrated potential for Psychrobacter sp. as a novel probiont to improve feed utilization, digestive enzymes and innate immunity in E. coioides.