微生物制剂对奥尼罗非鱼生长及消化酶活性的影响

选用192尾初始体重(34.500.25) g的健康奥尼罗非鱼(Oreochromis niloticusO. aureu), 研究在基础饲料中分别添加相同剂量(活菌含量为3.01011 cfu/kg饲料)的汉逊德巴利酵母、枯草芽孢杆菌和凝结芽孢杆菌对奥尼罗非鱼生长及消化酶活性的影响, 试验期56d。试验结果表明, 与对照组相比, 添加枯草芽孢杆菌和凝结芽孢杆菌, 奥尼罗非鱼的增重率分别提高12.27%和8.56%(P0.05), 饵料系数分别降低10.92%和8.18%(P0.05)。饲料干物质表观消化率分别提高10.54%和10.07%(P0.05), 蛋白质表观消化率分别提高4.18%和3.63%(P0.05)。添加枯草芽孢杆菌和凝结芽孢杆菌组, 奥尼罗非鱼肠道、肝胰脏和胃蛋白酶活性显著高于对照组和汉逊德巴利酵母组(P 0.05), 而添加三种微生物制剂对奥尼罗非鱼肠道、肝胰脏和胃的淀粉酶和脂肪酶没有显著影响(P0.05)。结果显示, 三种微生物制剂相互比较, 饲料中添加剂量为3.01011 cfu/kg饲料的枯草芽孢杆菌和凝结芽孢杆菌能显著提高肠道、肝胰脏和胃的蛋白酶活性, 满足最佳生长。       

枯草芽孢杆菌微生态制剂发酵研究进展

微生态制剂是饲用抗生素的绿色有效替代品。枯草芽孢杆菌在逆境中可形成抗逆性强的芽孢,在生产和应用过程中保持高活性,是一种高效的微生态制剂菌种。提高枯草芽孢杆菌活菌数及芽孢率是保证微生态制剂产品质量的关键。本文综述了枯草芽孢杆菌芽孢形成的分子生物学机制及影响芽孢形成的重要因素,进一步比较枯草芽孢杆菌微生态制剂不同发酵方式的特点,重点阐述了提高枯草芽孢杆菌有效生物量的工艺优化,最后介绍了枯草芽孢杆菌微生态制剂的应用,并对将来研究思路进行了讨论。     

饲料中添加枯草芽孢杆菌对青鱼生长、消化酶活性和鱼体组成的影响

用不加枯草芽孢杆菌的对照饲料和分别添加2×10⁶、2×10⁷、2×10⁸和5×10⁸ cfu/g枯草芽孢杆菌的4种实验饲料, 分别喂养初始均重为(4.16±0.03) g的5组三重复的青鱼鱼种8周。饲养试验在15只容积为80 L循环式水簇箱内进行, 水温控制为(25±0.5) ℃。结果表明: 饲料中添加枯草芽孢杆菌的实验组青鱼鱼体终重和鱼体增重比不添加的饲料组显著提高, 而饲料系数则显著降低(P<0.05), 不同枯草芽孢杆菌添加量对青鱼鱼体增重和饲料系数均无显著影响(P>0.05)。饲料中添加枯草芽孢杆菌对青鱼的成活率无显著影响(P>0.05)。青鱼摄食添加枯草芽孢杆菌的实验饲料后其肠蛋白酶活性比不添加组显著升高(P<0.05), 并随着枯草芽孢杆菌添加量的增加而显著上升(P<0.05), 但添加量到2×10⁷ cfu/g后继续增加则不再显著变化(P>0.05)。青鱼摄食添加枯草芽孢杆菌的实验饲料后, 其肠淀粉酶活性均比不添加组显著升高(P<0.05), 且随枯草芽孢杆菌添加量增大呈不断升高的趋势。青鱼摄食添加2×10⁷、2×10⁸和5×10⁸ cfu/g枯草芽孢杆菌的饲料后, 其肝胰脏蛋白酶活性显著高于摄食不添加或添加2×10⁶ cfu/g枯草芽孢杆菌的实验组(P<0.05)。青鱼摄食添加枯草芽孢杆菌的实验饲料后, 其肝胰脏淀粉酶、肝胰脏脂肪酶和肠脂肪酶的活性均与不添加的实验组无显著差异(P>0.05)。青鱼鱼种摄食添加枯草芽孢杆菌的饲料与摄食不添加枯草芽孢杆菌的饲料相比, 其全鱼营养组成均无显著差异(P>0.05)。因此, 在实验条件下, 饲料中添加2×10⁶—5×10⁸ cfu/g的枯草芽孢杆菌能促进青鱼鱼种的生长和降低饲料系数, 而对鱼体组成没有影响。     

凝结芽孢杆菌与酵母核苷酸对团头鲂生长和肠道的影响

目的 考察饲料中单独添加凝结芽孢杆菌或与酵母核苷酸联合添加,对团头鲂生长和其肠道的影响.方法 以团头鲂为试验对象随机分为3组,对照组饲喂基础饲料,A试验组饲喂“基础饲料+1 000 mg/kg凝结芽孢杆菌制剂”;B试验组饲喂“基础饲料+1 000 mg/kg凝结芽孢杆菌制剂+200 mg/kg酵母核苷酸”.饲喂56 d后,测定鱼体的各项生长指标、小肠绒毛高度、肠道菌群数量和消化酶活力.结果 与对照组比较,A组的特定生长率、蛋白质效率、肠道凝结芽孢杆菌数量和蛋白酶活力显著提高(P＜0.05),饲料系数和肠道大肠埃希菌数量显著降低(P＜0.05),但肠绒毛高度和肠道淀粉酶活力无明显增加(P＞0.05).与对照组和A组比较,B组的特定生长率、蛋白质效率、肠绒毛高度、肠道凝结芽孢杆菌数量、蛋白酶和淀粉酶活力均有显著提高(P＜0.05),饲料系数和肠道大肠埃希菌数量均有显著降低(P＜0.05).3组的摄食率没有明显改变(P＞0.05).A组和B组均不能提高肠道二糖酶活力(P＞0.05).结论 凝结芽孢杆菌与酵母核苷酸能协同促进团头鲂的生长和其肠道功能.     

微生态叶面肥促进小麦生长的效应

研究了稀释100倍的枯草芽孢杆菌、沼泽红假单胞菌、乳酸菌、假丝酵母菌单一菌种微生态叶面肥对小麦生长的效应。结果表明微生态叶面肥对小麦生长有促进作用,其活菌制剂对小麦的生长促进效应更为显著。其中,沼泽红假单胞菌有效的提高成穗率,较对照组增幅为10.2%;枯草芽孢杆菌对小麦的穗粒数和百粒重增加效果明显,较对照组的增幅分别为14.19%和17.01%;乳酸菌则有效的促进了小麦的分蘖及穗长,较对照组的增幅分别为16.24%和6.37%。而酵母菌的促进效果不明显。     

常见4种微生态制剂菌种产淀粉酶的比较

目的 探索常见4种微生态制剂菌种地衣芽胞杆菌、枯草芽胞杆菌、蜡样芽胞杆菌和纳豆芽胞杆菌产生淀粉酶的能力,以筛选和研制动物微生态制剂和饲料添加剂的使用菌种。方法 将各菌种接于淀粉酶试验培养基,培养后滴加稀碘溶液,观察透明圈,判定产酶能力。结果 地衣芽胞杆菌、枯草芽胞杆菌、蜡样芽胞杆菌和纳豆芽胞杆菌都能产生淀粉酶,以蜡样芽胞杆菌产生的淀粉酶较多。结论 4种常见芽胞杆菌产淀粉酶能力依次为：蜡样芽胞杆菌＞纳豆芽胞杆菌＞枯草芽胞杆菌＞地衣芽胞杆菌。     

益生菌ZD02的分离及其在对虾集约化精养中的应用

从养殖环境分离纯化芽孢杆菌6株,对其进行消化酶活检测,根据检测的酶活指标选择ZD02作为待试验菌株并进行鉴定和热稳定性检测。结果表明:ZD02被鉴定为枯草芽孢杆菌,具有耐受高温的特性,90℃水浴10min存活率93%,95℃水浴5min存活率93%,100℃水浴5min存活82%,95℃和100℃水浴2min不影响芽孢菌存活,在调质温度102℃条件下,制粒后芽孢菌存活率94%;将经过鉴定的枯草芽孢杆菌发酵培养后应用于凡纳滨对虾(Litopenaeus vannamei)的集约化精养殖,试验组A在基础饲料中添加0.3%的芽孢杆菌。试验组B保持配方成本不变的基础上添加0.3%的芽孢杆菌,结果显示,试验组的成活率显著高于对照组(P0.05),试验组A、B的产量分别比对照组提高了43.0%和36.7%(P0.05),产值分别提高35.4%和21.0%(P0.05),饲料系数比对照组分别降低了9.9%和8.7%(P0.05);换水耗电、水质调节剂、内服药物3项费用合计,A组和B组较对照组分别节省了21.3%和22.3%(P0.05);实验组之间各项指标差异不显著(P0.05);表明,在对虾的集约化精养过程中合理利用益生芽孢杆菌可以增产增收。     

复合微生态制剂对幼刺参营养成分和 酶活力的影响

目的研究复合微生态制剂对幼刺参体壁营养成分、几种消化酶和免疫酶活力的影响。方法在封闭式循环系统中投喂不同的微生态制剂进行30 d刺参养殖实验。结果投喂液态复合微生态制剂组和粉状复合微生态制剂组的刺参体壁的粗脂肪、总糖、粗蛋白含量最高,两组与对照组相比差异有统计学意义(Ps0.05)。添加微生态制剂的3个实验组刺参肠道蛋白酶、淀粉酶活力均比对照组高,且差异有统计学意义(Ps0.05)。实验组刺参的肠道、体壁、体液过氧化氢酶活力均比对照组高,其中投喂液态复合微生态制剂实验组活力最高。实验组刺参组织的酸性磷酸酶和碱性磷酸酶活力也明显高于对照组(与对照组相比差异有统计学意义,Ps0.05)。结论微生态制剂可以有效改善幼刺参体壁营养成分,促进机体消化活力,提高刺参免疫力。     

微生态制剂在水产养殖中的作用

微生态制剂的主要作用有 :净化水质、改善养殖水体环境 ;抑制病原微生物 ,提高机体免疫力 ;作为饲料添加剂 ,促进养殖生物生长 ;常用的微生物菌群主要有光合细菌、硝化细菌、芽孢杆菌等。综述了微生态制剂在水产养殖中的作用及常用微生物菌群。     

短短芽孢杆菌(Brevibacillus brevis)对鲤鱼养殖水质及其生长特性的影响

试验共设单因素4 水平, 分别对应水体中含0 mL、20 mL、60 mL、100 mL4 种体积短短芽孢杆菌的试验组。通过测定养殖水体COD、NH4+-N、DO、pH 等指标, 鲤鱼生长性能及消化酶活性等指标, 研究短短芽孢杆菌(Brevibacillus brevis)对鲤鱼养殖水体水质及鲤鱼生长特性的影响。结果显示, 短短芽孢杆菌能有效降低养殖水体中COD 和NH4 +-N 浓度(P<0.05), 维持DO 在较高水平(P<0.05), 保持pH 稳定, 改善养殖水质。试验组中鲤鱼的肥满度、增重率及特定生长率与对照组相比均显著提高(P<0.05), 鲤鱼肠道蛋白酶、淀粉酶和肝胰脏淀粉酶活性与对照组相比显著升高(P<0.05)。     

Effects of potential probiotic Bacillus subtilis T13 on growth, immunity and disease resistance against Vibrio splendidus infection in juvenile sea cucumber Apostichopus japonicus

A feeding experiment was conducted to determine influences of potential probiotic Bacillus subtilis T13 (isolated from intestine of healthy sea cucumbers) on growth, immunity and disease resistance against Vibrio splendidus infection in juvenile sea cucumbers Apostichopus japonicus. Animals were fed with diets containing B. subtilis T13 at 0, 10(5), 10(7) and 10(9) CFU/g for 30 days, respectively. At the end of the growth trial, fifteen sea cucumbers from each aquarium were sampled for immune indices measurement. Then twenty sea cucumbers from each replicate were challenged with V. splendidus. Results showed that administration of B. subtilis T13 had significant effect on the specific growth rates (SGR) of sea cucumbers (P < 0.05). Phagocytosis, respiratory burst activity and total nitric oxide synthase (T-NOS) activity were significantly improved in coelomocytes of sea cucumbers fed with T13 at 10(9) CFU/g diet (P < 0.05). The highest values of the total coelomocytes counts (TCC) and superoxide dismutase (SOD) activity were found in sea cucumbers fed diet containing T13 at 10(9) CFU/g. The cumulative mortality after V. splendidus challenge decreased significantly in sea cucumbers fed with T13 at dose of 10(9) CFU/g (P < 0.05). The present study confirmed the potential beneficial effects of B. subtilis T13 as dietary probiotic in juvenile A. japonicus.     

高效降解蛋白枯草芽孢杆菌的筛选及促建鲤生长的研究

【目的】从建鲤肠道分离并优选出了一株对蛋白具有强效降解力的菌株,探究其临床生产效果。【方法】使用牛奶平板进行筛选,经常规细菌学鉴定和16S rDNA序列分析确定A7菌株为枯草芽孢杆菌,通过临床饲喂试验探究其对建鲤促生长作用。【结果】A7菌株在牛奶平板上的水解圈直径可达27.5 mm,其最佳固体发酵条件为温度28°C、接种量5%(1.2×109 CFU/mL)、料水比1.0:1.2和发酵时间72 h。临床饲喂试验结果表明,饵料中添加0.5%、1.0%和1.5%的A7株菌粉均能促建鲤生长。其中,添加量为1.0%的试验组,鱼体增重率和蛋白利用率最高,饵料系数最低,与产品对照组和空白对照组相比,均达到了显著性差异(P0.05)。此外,随A7株菌粉添菌量的增加,各试验组肝胰脏和肠道的蛋白酶及淀粉酶活力出现先增大后减小趋势,与产品对照组和空白对照组相比,添加1.0%的试验组增幅最大(P0.05)。研究发现,试验组鱼肌肉中粗蛋白和粗脂肪含量较空白对照组也有明显的增减趋势(P0.05)。【结论】饵料中添加A7株菌粉,可有效促进鱼的生长,降低饵料系数,提高肝胰脏和肠道的蛋白酶及淀粉酶活力,并提高鱼肌肉中的蛋白含量和降低脂肪含量。     

学生食堂餐厨垃圾中淀粉的生物利用菌株筛选

目的探索地衣芽胞杆菌、枯草芽胞杆菌和蜡样芽胞杆菌分解大学生食堂厨余中淀粉的能力,以筛选和研制餐厨垃圾生物降解的使用菌种。方法将各菌种接于淀粉酶试验培养基,培养后滴加碘溶液,观察透明圈,判定产淀粉酶能力;收集大学生食堂的厨余,观察三种细菌在不同接种量（5%、10%、15%、20%、25%）、不同接种时间（24 h、48 h、72 h）及不同菌株配伍方式下发酵淀粉的能力。结果地衣芽胞杆菌、枯草芽胞杆菌、蜡样芽胞杆菌都能产生淀粉酶,以地衣芽胞杆菌产生的淀粉酶较多,其次为枯草芽胞杆菌,蜡样芽胞杆菌较少,三种细菌分解厨余中淀粉的最佳接种量都为15%-20%,最佳发酵时间为48 h,枯草芽胞杆菌和地衣芽胞杆菌各7.5%的接种量混合配伍发酵效果最佳。结论可采用枯草芽胞杆菌和地衣芽胞杆菌各7.5%的接种量混合配伍发酵学生食堂的厨余中的淀粉。     

Distribution and characteristics of Bacillus bacteria associated with hydrobionts and the waters of the Peter the Great Bay, Sea of Japan

Bacilli of the species Bacillus subtilis, B. pumilus, B. mycoides, B. marinus and B. licheniformis (a total of 53 strains) were isolated from 15 invertebrate species and the water of the Vostok Bay, Peter the Great Bay, Sea of Japan. Bacilli were most often isolated from bivalves (22.7%) and sea cucumbers (18.9%); they occurred less frequently in sea urchins and starfish (13.2 and 7.5%, respectively). Most of bacilli strains were isolated from invertebrates inhabiting silted sediments. No Bacillus spp. strains were isolated from invertebrates inhabiting stony and sandy environments. The species diversity of bacilli isolated from marine objects under study was low. Almost all bacterial isolates were resistant to lincomycin. Unlike B. pumilus, B. subtilis isolates were mostly resistant to benzylpenicillin and ampicillin. Antibiotic sensitivity of B. licheniformis strains was variable (two strains were resistant to benzylpenicillin and oxacillin, while one was sensitive). A significant fraction of isolated bacilli contained pigments. Pigmented strains were more often isolated from seawater samples, while colorless ones predominated within hydrobionts. B. subtilis colonies had the broadest range of colors. In the Bacillus strains obtained, DNase, RNase, phosphatase, elastolytic, chitinase, and agarolytic activity was detected. Bacilli strains with hydrolytic activity occurred in invertebrates more often than in seawater.     

Extracellular amylase production by cassava-fermenting bacteria

Summary Fermentation of cassava tubers was accompanied by a gradual decrease in pH, increased amylase activity in the steep liquor, and increased microbial load and lactic acid concentration. Amylase-producing bacterial strains associated with cassava fermentation were isolated and identified asBacillus subtilis, Bacillus licheniformis andBacillus cereus. The pH optima for the partially purified enzymes of these organisms were 7.0, 5.5 and 7.5, whilst their temperature optima were 30, 37 and 80°C. There was no significant difference in amylase activities when starch, dextrin, amylopectin, glucose and maltose were used as growth substrates.     

Biocontrol and other beneficial activities of Bacillus subtilis isolated from cowdung microflora

Bacillus subtilis strains isolated from cowdung (CD) had several beneficial attributes, which included biocontrol, plant growth promotion, sulphur (S) oxidation, phosphorus (P) solubilization and production of industrially important enzymes (amylase and cellulase). The B. subtilis strains from CD inhibited the in vitro growth of fungi, Fusarium oxysporum (25-34%) and Botryodiplodia theobromae (100%), isolated from the postharvest rots of yam (Dioscorea rotundata) tubers. Other than biocontrol, B. subtilis strains were able to promote root elongation in seedlings of Cicer arietinum up to 70-74% as compared to untreated seeds (control). B. subtilis strains had also the ability to oxidize elemental S to sulphate (2-15microgml(-1)) and showed distinct P-solubilization activity in vitro. In addition, the cultures showed cellulase activity in carboxy methyl cellulose medium (1.5-1.8mg of reducing sugar24h(-1)ml(-1)) and amylase activity in vitro.     

人参茎叶提取物对刺参生长、免疫及抗病力的影响

以初始体重为(4.910.17) g的刺参(Apostichopus japonicas)为研究对象,研究投喂不同剂量人参茎叶提取物对刺参生长、免疫及抗病力的影响,探讨人参茎叶提取物在刺参养殖中的应用。在每千克基础饲料中分别添加10、20、40和80 g人参茎叶提取物作为实验组,并以基础饲料为对照进行为期30d的饲喂养殖。结果显示,投喂人参茎叶提取物对刺参的生长并无显著影响,刺参特定生长率不随人参茎叶提取物添加量的增加而提高。当人参茎叶提取物添加量为80 g/kg时,刺参体腔细胞总数、呼吸暴发强度、细胞吞噬活性、超氧化物歧化酶活力、总一氧化氮合酶活力均显著高于对照组(P0.05);当人参茎叶提取物添加量为40 g/kg时,细胞吞噬活性也显著高于对照组(P0.05),但其余免疫指标与对照组相比无显著差异(P0.05);当人参茎叶提取物的添加量不大于20 g/kg时,各项免疫指标与对照组之间均无显著差异(P0.05)。攻毒实验表明, 20、40和80 g/kg人参茎叶提取物组的健康率要高于对照组,其中80 g/kg人参茎叶提取物组健康率为80.56%要显著高于对照组和10 g/kg人参茎叶提取物组(P0.05)。人参茎叶提取物可作为免疫增强剂应用于刺参的养殖中来提高刺参的免疫和抗病能力。       

表达淀粉酶、糖化酶葡糖酸醋杆菌工程菌的构建

采用基因融合技术,将葡糖酸醋杆菌Gluconacetobacter hansenii ATCC23769分泌蛋白CMCax的信号肽序列分别与来源于枯草芽胞杆菌的淀粉酶基因、黑曲霉的糖化酶基因融合构建融合蛋白,连入能在G.hansenii ATCC23769自主复制的载体pbs-H1S中,电击转入G.hansenii ATCC23769,构建能内源表达淀粉酶、糖化酶,以及淀粉酶-糖化酶的葡糖酸醋杆菌。淀粉平板透明圈检测结果和DNS测酶活结果显示,构建的3种工程菌能成功表达并分泌淀粉酶和糖化酶。