饲喂枯草芽孢对草鱼粪便、养殖水体中芽孢数量和水质的影响

分别在饲料中添加不同含量的枯草(Bacillus subtilis)芽孢(A组实测芽孢数量为2.6×107 cfu/g,B组为4.4×106 cfu/g,空白组为0),饲喂静水水族箱中100 g左右的草鱼(Ctenopharyngodon idellus)10 d,测定饲喂枯草芽孢后草鱼粪便、养殖水体中芽孢数量和水质的变化。结果表明:(1)9 d内,水体中芽孢数量A组和B组在前6 d时不断增加,在6 d时达到稳定,其中A组和B组稳定时的含量分别为2.22×106 cfu/g和3.98×105 cfu/g。(2)10 d内,草鱼粪便干物质中的芽孢数量与饲料中的枯草芽孢含量相比,A组和B组粪便中的枯草芽孢损失率分别为89.76%～90.71%和83.83%～86.84%。(3)草鱼饲喂枯草芽孢后,A组和B组粪便中排出的枯草芽孢对水体中亚硝酸盐和氨氮有降低的趋势,但各组之间差异不显著(P>0.05),对化学需氧量(COD)没有影响。     

草鱼肠道芽孢杆菌的鉴定及产酶能力分析

从100 g左右的健康草鱼(Ctenopharyngodon idellus)肠道中分离出3株疑似芽孢杆菌(Bacillus)菌株(G1、G2、G3),并通过对其进行生理生化特征、16S rDNA全序列和产酶能力的分析,筛选出1株高产酶能力的益生菌株。结果显示:G1、G2和G3菌株与枯草芽孢杆菌在16S rDNA序列相似性≥99%的水平上聚为同一分支,结合形态观察和生理生化特征,最终鉴定G1、G2和G3菌株均为枯草芽孢杆菌(Bacillus subtilis);以水解圈直径/菌落直径比值评定了G1、G2和G3菌株的产纤维素酶、淀粉酶和蛋白酶的能力,其中G1菌株产纤维素酶、淀粉酶和蛋白酶能力(比值分别为2.9、2.2和3.3)均高于G2、G3菌株,具有作为益生菌的潜力。     

饲料中添加复合芽孢杆菌对草鱼消化道酶活性及肠道菌群的影响

用含复合芽孢杆菌(105cfu/g饲料,枯草芽孢杆菌和解淀粉芽孢杆菌以1∶1比例混合)的基础日粮,按体重3%的日投饵量,饲喂体重(51.37±0.58)g的草鱼(Ctenopharyngodon idellus),饲养实验45 d。结果表明:与对照组相比,处理组肠道内容物胰蛋白酶活性提高了79.46%(P<0.01);肝胰腺脂肪酶活性提高了16.82%(P<0.01),肝胰腺淀粉酶和胰蛋白酶活性无显著差异(P>0.05)。肠道菌群数量分析显示,处理组肠道芽孢杆菌数均显著提高(P<0.01),弧菌和大肠杆菌数显著降低(P<0.01)。结果提示,饵料中添加复合芽孢杆菌能够改善草鱼肠道菌群组成,并提高消化道特定消化酶活性。     

饲喂枯草芽孢杆菌对银鲫等水生动物肠道菌群及消化酶活性的影响

应用产酶(蛋白酶、脂肪酶和淀粉酶)能力较强的枯草芽孢杆菌B115制成微生物添加剂，饲料中添加量为2‰。经35d投喂试验，测定翘嘴红鲐、银鲫、日本沼虾胃肠道消化酶活性、肠道微生物菌群和生长的变化。结果表明，添加枯草芽孢杆菌B115，可显著提高鱼虾肠道消化酶的活性。从银鲫整个肠道的平均值看，试验组比对照组的蛋白酶、脂肪酶和淀粉酶活性分别提高19．8％、22．7％和11．03％；从翘嘴红鲐整个肠道的平均值看，试验组比对照组分别提高13．1％、15．8％和13．9％。日本沼虾试验组胃的蛋白酶、脂肪酶和淀粉酶活性分别高于对照组13．69％、10．05％和10．44％，试验组肠的蛋白酶、脂肪酶和淀粉酶活性分别高于对照组8、8％、15．5％和8．43％。微生物添加剂组对鱼虾的生长有促进作用，同时能增加肠道中有益菌群如乳杆菌和双歧杆菌，对气单胞菌、肠杆菌和肠球菌的量基本没有影响。     

枯草芽孢杆菌对杂交鲟幼鱼生长性能、消化酶活性及非特异性免疫的影响

在杂交鲟(Acipenser baeri ♂×Acipenserschrenkii♀)幼鱼日粮中添加不同质量分数(0、0.05％、0.1％、0.25％、0.5％)的枯草芽孢杆菌(Bacillussubtilis)粉剂,分别在培育实验的第15天和第30天采集样本,分析其对幼鱼摄食、生长、肠道消化酶活性及非特异性免疫指标的影响.实验结果:饲料中添加0.25％枯草芽孢杆菌粉剂时,杂交鲟生长最好,增重率和特定生长率较对照组分别提高了54.31％(P＜0.05)和28.61％(P＜0.05),饵料系数降低了36.02％(P＜o.05),也显著低于其他各实验组(P＜0.05).各实验组肠道蛋白酶、淀粉酶、脂肪酶活性均高于对照组;实验第15天时各种消化酶活性均大于第30天,其中第15天添加0.25％组的肠道蛋白酶、淀粉酶和脂肪酶活性最高,显著高于对照组(P＜0.05),且该实验组的血清溶菌酶(LSZ)、碱性磷酸酶(AKP)、总抗氧化能力(T-AOC)及超氧化物歧化酶(SOD)均显著高于对照组和其他实验组(P＜0.05).研究表明,饲料中添加枯草芽孢杆菌可提高杂交鲟幼鱼的生长性能、肠道的消化酶活性及血清非特异性免疫能力,适宜添加水平为0.25％,添加的适宜持续时间为15d左右.     

枯草芽孢杆菌的培养条件及对水质的净化作用

研究了枯草芽孢杆菌(Bacillus subtilis Cohn,1872)(菌株编号:LB-B3)的培养条件及其在净化对虾养殖池水样方面的效果。实验分别设置了9个不同的pH值梯度(pH 2～10)和6个不同接种量梯度(0.3%、0.5%、1%、3%、5%和7%),以吸光值(OD)为生长指标,进行了枯草芽孢杆菌培养条件的优化实验;同时又设置了5个不同接种浓度(0 CFU/mL、5.2×104CFU/mL、1.04×105CFU/mL、1.56×105CFU/mL和2.08×105CFU/mL)接种待处理水样,测定了96 h内化学耗氧量、亚硝酸氮、氨态氮和溶氧等4个水质指标的变化情况。结果表明:pH=7.0、接种量为7.0%时,OD值最大;枯草芽孢杆菌能显著净化水质,但使用后会暂时性增加耗氧。     

枯草芽孢杆菌对克氏原螯虾免疫机能的影响

在饲料中分别添加0、1×109、3×109和5×109cfu/kg枯草芽孢杆菌(Bacillus subtilis),测定投喂相应饲料后第1、4、7、14、21、28天以及停止投喂枯草芽孢杆菌后的第7、14天克氏原螯虾(Procam barus clarkii)的血淋巴吞噬活性、血清溶菌活力、抗菌活力以及酚氧化酶活力。结果显示:实验组(含枯草芽孢杆菌的饲料组)的吞噬活性于投喂枯草芽孢杆菌后第7天开始显著高于对照组(P<0.05);实验组的溶菌活力、抗菌活力明显高于对照组(P<0.05),并在停药后仍维持较高水平(P<0.05);此外,酚氧化酶活力也有所增加。本研究表明,枯草芽孢杆菌对克氏原螯虾的免疫机能有促进作用。     

酸化剂对草鱼生长和非特异性免疫能力的影响

为考察不同类型酸化剂对草鱼生长及生理机能的影响,在基础饲料(对照组)中分别添加0.21％磷酸、柠檬酸、乳酸和复合酸,饲养初始体重为(10.33±0.03)g草鱼(Ctenopharyngodon idellus)8周.结果表明:(1)与对照组相比,饲料中添加0.21％柠檬酸、乳酸和复合酸均显著提高草鱼的增重率、特定生长...     

博斯腾湖草鱼生长特征的研究

根据2014年8月采集的33尾标本对博斯腾湖草鱼(Ctenopharyngodon idellus)的生长特征进行研究。以鳞片为年龄和生长退算材料,研究结果显示:博斯腾湖草鱼体重与体长表现为异速生长,关系式为W=0.0444SL~(2.8322)。用退算体长拟合的von Bertalanffy生长参数为:SL_8=103.7664 cm,k=0.1385/y,t_0=-0.0542 y。体重生长参数为:W_8=22.7674 kg,k=0.1385/y,t_0=0.0027 y。生长特征指数为3.16;生长拐点年龄为7.5龄,拐点时体长和体重分别为67.3 cm和6.6 kg。与1979年和2000年种群相比,受饵料供给不足的制约,博斯腾湖草鱼生长速度明显下降。草鱼增殖管理措施的制定应以恢复沉水植物资源为目标,具体措施包括:减少草鱼的投放量,草鱼最小捕捞规格降至3 kg,提高草鱼的捕捞强度和捕捞量。     

光合细菌和枯草芽孢杆菌对养殖水质的净化作用

以光合细菌(Photosynthetic bacteria,P)和枯草芽孢杆菌(Bacillus subtilis,B)为实验菌种,研究两者最佳浓度配比的复合菌组对淡水养殖水质的净化作用。试验设置1个对照组(CK)和5个复合菌组(PB1、PB2、PB3、PB4、PB5),5个复合菌组浓度配比分别为(2.0×105+1.5×105)CFU/m L、(2.0×105+3.0×105)CFU/m L、(2.0×105+4.5×105)CFU/m L、(4.0×105+1.5×105)CFU/m L、(6.0×105+1.5×105)CFU/m L,分析各试验组的化学需氧量(COD)、氨氮(NH3-N)、溶解氧(DO)、p H等水质指标。结果显示:复合菌能够明显去除水体CODMn,PB2组去除率最高达,44.98%;能有效增加DO,PB2组增氧率最高,为27.9%;能明显去除氨氮,PB2组去除率最高达78%;并且能稳定p H值在8.6左右,5个复合菌组差异不显著(P0.01)。复合菌发挥最佳净化能力的时间约在6~8 d。结果表明,复合菌最佳浓度配比为(2.0×105+3.0×105)CFU/m L,该浓度组较对照组和其他试验组能够显著净化淡水养殖水质,有效改善养殖环境。     

不同蛋白能量比饲料对草鱼幼鱼消化酶活性的影响

以鱼粉和豆粕为蛋白源,鱼油和豆油等比例混合油为脂肪源,共配制4个蛋白水平(20%、25%、30%、35%),每一蛋白水平设4个脂肪水平(4.5%、8%、11.5%、15%),共16组饲料。每组设3个重复,在水温23~29℃下,连续投喂初始体重为(16.84±0.28)g的草鱼(Ctenopharyngodon idellus)10周,研究不同蛋白能量比饲料对草鱼幼鱼消化酶活性的影响。结果显示:肠道蛋白酶的活性随着饲料蛋白含量的升高而显著升高(P<0.05),饲料蛋白水平在20%~30%时,饲料中的脂肪水平对蛋白酶活性影响不显著(P>0.05),蛋白水平在35%时,高脂肪抑制了蛋白酶活性,肝胰脏的蛋白酶变化规律类似于肠道;肠道和肝胰脏淀粉酶活性在饲料蛋白含量相同的情况下,随着脂肪含量的升高呈现显著的下降趋势(P<0.05),但饲料中蛋白(30%,35%)和脂肪(8%,11.5%,15%)含量较高时,淀粉酶活性较稳定;肠道和肝胰脏的脂肪酶活性在饲料蛋白含量相同的情况下,随着脂肪含量的升高,呈显著上升的趋势(P<0.05),但在蛋白含量(30%,35%)和脂肪含量(8%、11.5%,15%)的6个试验组之间差异不显著(P>0.05);在脂肪含量分别为4.5%、8%和11.5%时,饲料的蛋白含量对肠道的脂肪酶活性无显著性影响(P>0.05),当脂肪含量为15%时,随着蛋白含量的升高,肠道的脂肪酶活性显著降低(P<0.05)。结果表明,草鱼饲料中蛋白含量为30%和脂肪为8%(即蛋白能量比21.7 mg/kJ)的试验组草鱼获得较高的消化酶活性。     

饲料蛋白能量比对草鱼幼鱼生长性能、蛋白利用和体成分的影响

以鱼粉和豆粕为蛋白源,鱼油和豆油等比例混合油为脂肪源,采用4×4因子实验来确定草鱼(Ctenopharyngodon idellus)幼鱼饲料中的合适蛋白能量比(P/E),其中饲料蛋白水平分别为20%、25%、30%、35%,能量水平为12.5、13.7、15、16.2 k J/g,饲料蛋白能量比的范围为12.1~27.3 mg/k J,共16组饲料,每组设3个重复,连续投喂初体质量(16.85±0.29)g的草鱼幼鱼10周,研究饲料蛋白能量比对草鱼幼鱼生长性能、蛋白利用和体成分的影响。结果表明:(1)最高末体质量和特定生长率出现在P/E为18.3 mg/k J组,而P/E为16.0 mg/k J组的饲料系数最高,蛋白质效率和蛋白沉积率最高组分别为P/E 12.10 mg/k J组和15.20 mg/k J组。(2)P/E为27.30 mg/k J组的能量沉积率和总氮排泄率最大。(3)在同一饲料蛋白水平下,鱼体的脂肪含量随饲料能量水平升高而升高;在同一饲料能量水平下,鱼体的蛋白含量随饲料蛋白含量升高而升高。对生长、蛋白利用、体成分的实验结果进行综合分析,草鱼幼鱼饲料的最适蛋白含量为30%,最适P/E为19.5 mg/k J。     

Effects of dietary selenium on growth performance and oxidative stress in juvenile grass carp Ctenopharyngodon idellus

An 8‐week feeding trial was conducted to investigate the effect of dietary selenium (Se) on feed intake, weight gain and antioxidant activity in juvenile grass carp (11.2 ± 0.03 g). Six Se levels (0.13, 0.41, 0.56, 1.12, 2.18 and 4.31 mg/kg) of semi‐purified diets were assayed in triplicate. The maximum weight gain, specific growth rate and feed intake were obtained in fish fed with 1.12 mg Se/kg diet. Hepatic glutathione peroxidase activity was markedly increased when dietary Se ≤1.12 mg/kg diet and reached a plateau when dietary Se ≥1.12 mg/kg diet. Hepatic superoxide dismutase and serum catalase activities in juvenile grass carp fed with 0.56, 1.12 and 2.18 mg Se/kg diets were all significantly higher than those in the other groups. The malondialdehyde content in liver and serum was firstly decreased and then increased with increasing dietary Se content, and the lowest content was observed in fish fed with 1.12 mg Se/kg diet. With the increase in Se level, the activities of serum alanine aminotransferase and aspartate aminotransferase were reduced. In addition, serum alkaline phosphatase activity and albumin content were highest in fish fed with 1.12 mg Se/kg diet. This study indicated that both the Se deficiency and excess of Se caused negative effect on the oxidative stress in juvenile grass carp and suggested that the health‐giving concentration of dietary inorganic Se was 1.12 mg/kg diet. Moreover, based on the broken‐line regression analysis of weight gain, the optimal concentration of dietary inorganic Se was 0.83 mg/kg for juvenile grass carp.     

Effect of dietary inclusion of cottonseed meal on growth performance and physiological and immune responses in juvenile grass carp,Ctenopharyngodon idellus

An 8‐week feeding trial was conducted to evaluate the effect of replacement of dietary fishmeal with cottonseed meal (CSM) on the growth performance and health status of juvenile grass carp, Ctenopharyngodon idellus (6.67 g). Six isonitrogenous and isoenergetic diets were formulated with different CSM levels (0, 121, 241, 362, 482 and 603 g/kg diet). The first limiting amino acid was methionine, which decreased with an increase in dietary CSM from 361 to 538 g/kg. When the dietary CSM content was lower than 241 g/kg, the limiting amino acid was histidine. Thermal growth coefficient (TGC) and feed efficiency first increased and then decreased (p < 0.05), which was consistent with the change in the limiting amino acid. The feeding rate increased with a rise in the dietary CSM level (p < 0.05). The activity of digestive enzymes except trypsin decreased significantly (p < 0.05), serum aminotransferase activity and triglycerides increased, serum cholesterol decreased (p < 0.05). When dietary CSM was raised to 362 g/kg, intestinal villi and hepatopancreas tissue were injured. When the dietary CSM was 603 g/kg, alternative complement pathway haemolytic activity was inhibited (p < 0.05). The gossypol level in dorsal muscles from each treatment was below the detection limit. Based on the quadratic regression analysis of the TGC, the optimal dietary CSM level was 175.9 g/kg, and the maximum could be up to 351.8 g/kg and still result in the same TGC as that of control group.     

Effect of replacing soybean meal with cottonseed meal on growth, hematology, antioxidant enzymes activity and expression for juvenile grass carp, Ctenopharyngodon idellus

An 8-week feeding trial was conducted to evaluate the effect of replacing soybean meal (SBM) with cottonseed meal (CSM) on growth and health of grass carp. Four isonitrogenous diets containing 0, 16.64, 32.73 and 48.94% of CSM, respectively, as replacements of 0, 35, 68 and 100% of SBM were fed to fish (initial body weight 7.14 ± 0.75 g/fish) in triplicate aquaria twice daily. The results indicated that fish fed diet containing 16.64% CSM as a replacement of 35% of SBM was not affected in weight gain (WG), feed efficiency ratio (FER) and feed conversion ratio (FCR) (P>0.05), while fish fed diets containing higher level of dietary CSM (32.73 and 48.94%) significantly decreased WGand PER and significantly increased FCR (P<0.05). Fish fed diets containing 16.64% of CSM had significantly increased hematocrit (Ht) and hemoglobin (Hb) values compared with fish fed with other diets (P<0.05). The activity of catalase (CAT), glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD), gene expression levels of GSH-Px and CAT, and content of malondialdehyde (MDA) were significantly lower for fish fed diets containing 16.64% CSM compared with fish fed other diets (P<0.05). These results showed 16.64% CSM could be used to replace 35% SBM in the diets of juvenile grass carp and without health impact.     

Effects of dietary Bacillus pumilus on growth performance,innate immunity and digestive enzymes of giant freshwater prawns (Macrobrachium rosenbergii)

The present study was conducted to investigate the effects of Bacillus pumilus on the growth performance, innate immunity and digestive enzymes of the giant freshwater prawn, Macrobrachium rosenbergii. Prawns (mean weight: 1.81 ± 0.01 g) were fed different levels of B. pumilus at doses of 0, 1 × 10⁷, 1 × 10⁸ and 1 × 10⁹ CFU/g to produce one control group and three experimental groups (G1, G2 and G3), respectively. After a 60‐day experimental feeding period, the results showed that the final weight, weight gain rate (WGR) and specific growth rate (SGR) were significantly higher in experimental groups than in the control (p < 0.05), whereas these improvements did not differ significantly (p > 0.05) among experimental groups. The improvements in phagocytic activity, respiratory burst (RBs) activity and the activity of catalase (CAT), nitric oxide synthase (NOS) and acid phosphatase (ACP) were significantly higher in G2 and G3 than in the control and G1 (p < 0.05). Further, the activity of CAT, NOS and ACP significantly increased with an increase in the dietary level of B. pumilus (p < 0.05). The activity of phenoloxidase (PO) in G3 was significantly higher than that in the control (p < 0.05). The activity of amylase was significantly higher in the groups G2 and G3 than that in the control group. G3 improved the activity of protease compared with the control (p < 0.05). In conclusion, the results of this study indicated that the potential use of B. pumilusat a dose of 1 × 10⁸ CFU/g feed could improve the growth, immunity and digestive enzymes of M. rosenbergii.     

Effects of dietary lipid level on growth performance,body composition and digestive enzymes activity of juvenile sea cucumber,Apostichopus japonicus

A feeding trial aimed to determine the effects of dietary lipid level on growth performance, body composition and digestive enzymes activity of juvenile sea cucumber, Apostichopus japonicus. Diets with six crude lipid levels (1.9, 13.8, 29.1, 43.6, 59.6 and 71.6 g kg^?1) were fed to sea cucumbers (initial weights 0.65 ± 0.01 g) at a density of 30 juveniles, once a day. After 60 days, body weight gain (BWG), specific growth rate (SGR), feed intake (FI) and protein efficiency ratio (PER) decreased with increasing dietary lipid levels. The sea cucumbers fed 1.9 g kg^?1 crude lipid showed significantly higher (P < 0.05) BWG than those of the sea cucumbers fed 59.6 and 71.6 g kg^?1 crude lipid. Intestinal protease and lipase activities generally increased with increasing dietary lipid levels. Eicosapentaenoic acid (EPA) content of body walls generally increased with increasing dietary lipid levels. Docosahexaenoic acid (DHA) content of body walls reached the maximum value at a dietary lipid level of 13.8 g kg^?1. N‐3 highly unsaturated fatty acid content followed the same pattern of DHA. According to the growth performance and body composition of sea cucumbers, it can be indicated that the optimum dietary lipid level for juvenile sea cucumbers is between 1.9 and 13.8 g kg^?1.