芽孢杆菌在水产养殖中的应用及产品现状

芽孢杆菌(Bacillus)是细菌的一科,生命力强,分布广,常见种类包括芽孢杆菌属、芽孢乳杆菌属、梭菌属等,目前可利用的芽孢杆菌有枯草芽孢杆菌、凝结芽孢杆菌、苏云金芽孢杆菌、地衣芽孢杆菌、蜡样芽孢杆菌等.在水产养殖中应用的主要是枯草芽孢杆菌,其可以直接应用于养殖水体或添加到饲料中使用,而最广泛应用的是作为水质改良剂.     

养虾如何选用菌制剂

养虾用的菌制剂首选芽孢杆菌，其次是光合细菌以及乳酸菌产品。芽孢杆菌一般产品为枯草芽孢杆菌、地衣芽孢杆菌、纳豆芽孢杆菌、腊状芽孢杆菌，是所知极少数可以分解大分子有机质如残饵粪便的菌种。     

投加微生物制剂对对虾养殖池总氨氮与亚硝酸氮的影响

监测并分析了单独或组合加入光合细菌、益生菌(酵母与乳杆菌)和芽孢杆菌后,对虾养殖池凌晨水体中氨氮和亚硝酸氮的变化。试验分为6组,即3个单独添加组(益生菌组、芽孢杆菌组、光合细菌组)和3个组合添加组(光合细菌与芽孢杆菌组、益生菌与芽孢杆菌组、光合细菌与益生菌组)。结果表明,添加微生物制剂后,益生菌组、芽孢杆菌组、光合细菌与芽孢杆菌组、益生菌与芽孢杆菌组池塘水体中总氨氮水平提高了41%~99.8%;而光合细菌组、光合细菌与益生菌组无显著性变化,但亚硝酸氮水平有所升高(107%~210%)。单独添加组水体中总氨氮水平的变化强弱顺序为芽孢杆菌>益生菌>光合细菌,组合添加组为芽孢杆菌与益生菌组>芽孢杆菌与光合细菌组>益生菌与光合细菌组。试验期间各组养殖池中总氨氮浓度为0.51~1.94 mg/L、亚硝酸氮水平为0.016~0.096 mg/L,均在安全浓度以下,说明添加微生物制剂未引起池塘氨氮和亚硝酸氮的毒性问题。     

关于芽孢杆菌(Bacillus)应用概述

芽孢杆菌属拉丁学名(Bacillus Cohn,1872),细胞呈直杆状,0.5～2.5μm×1.2～10μm,常以成对或链状排列,具圆端或方端。革兰氏染色阳性菌。产生芽孢,需氧或兼性厌氧。大多数有动力,无荚膜,多数溶血,通常过氧化氢酶阳性。包括对人和动物致病的炭疽芽孢杆菌,可引起食物中毒的蜡状芽孢杆菌,非致病性的枯草芽孢杆菌、蕈状芽孢杆菌、多粘芽孢杆菌等。其广泛应用于水产上、农业、科学研究、工业、医药卫生。     

针对一株地衣芽孢杆菌部分耐受性的探讨

<正>目前用作益生菌的芽孢菌主要有枯草芽孢杆菌、地衣芽孢杆菌和蜡样芽孢杆菌,它们分布广,存在于土壤、水、空气以及动物肠道等处,繁殖快,生命力强。有益芽孢杆菌可以在条件不利的情况下形成休眠体芽孢,因此在饲料的加工制粒、通过动物胃酸环境等过程中的稳定性相对较高。这些休眠体芽孢进入动物体内,会在肠道内发芽而成为具有活性的营养     

两种芽孢杆菌体外体内拮抗气单胞菌的试验

将蜡样芽孢杆菌Y1和巨大芽孢杆菌BM1259混合培养,结合平板活菌计数、抑菌活性测定及体内拮抗试验,研究其对水产动物气单胞菌的抑菌效果。体外拮抗试验结果表明,这两种芽孢杆菌均能明显抑制嗜水气单胞菌NQ的增殖,抑制率分别为53.74%和56.12%。不同温度下的抑菌活性试验显示,2种芽孢杆菌的抑菌活性温度分别为32℃和29℃。体内拮抗试验结果表明,巨大芽孢杆菌能显著降低异育银鲫肠道气单胞菌的数量。提示巨大芽孢杆菌可作为潜在的水产专用微生态制剂。     

5种芽孢杆菌对水体中无机磷的去除效果

在蒸馏水+KH₂PO₄的环境条件下进行了地衣芽孢杆菌(Bacillus licheniformis)、凝结芽孢杆菌(B.coagulans)、枯草芽孢杆菌(B.subtilis)、解淀粉芽孢杆菌(B.amyloliquefaciens)和胶冻样类芽孢杆菌(Paenibacillus mucilaginosus)对水中无机磷的去除效果研究，结果表明，地衣芽孢杆菌、凝结芽孢杆菌和枯草芽孢杆菌基本不具备除磷能力，解淀粉芽孢杆菌和胶冻样类芽孢杆菌有一定除磷能力且除磷效果相近，均是从10⁴ CFU/mL开始随着菌液浓度的增加，除磷效果逐步提高，在10⁷ CFU/mL浓度组达到最佳除磷效果；同时除磷效果与水力停留期间随时间的演化规律相似，都是在处理6 h时基本达到最佳除磷效果，而6～8 h期间除磷效果明显降低。     

枯草芽孢杆菌的分离及其净化水质的研究

对水产养殖水体及底泥中的芽孢杆菌进行分离,得到一株枯草芽孢杆菌,并研究了枯草芽孢杆菌对养殖凡纳滨对虾池水质的净化作用。试验池中枯草芽孢杆菌密度4×109个/L,间隔7 d后再以相同的剂量施放1次,于施放菌液后的第1、3、5、7、9、11天上午采集水样。试验池中COD、亚硝酸盐、H2S比对照池显著降低,总碱度显著上升,表明枯草芽孢杆菌对养殖凡纳滨对虾池水质具有明显的净化作用。     

益生菌在凡纳滨对虾育苗中的应用

实验研究了在凡纳滨对虾育苗中添加益生菌(芽孢杆菌、光合细菌或芽孢杆菌 光合细菌)对水体中pH值、H2S、COD、NH 4N、NO-2N和幼体成活率的影响。结果表明:光合细菌对提高水体的pH值有一定作用,而芽孢杆菌对pH值无显著影响;益生菌能显著降低水体中的H2S、NH 4N和NO-2N含量,其中光合细菌去除H2S和NO-2N的效果略优于芽孢杆菌,而芽孢杆菌去除NH 4N的能力比光合细菌强;实验中COD均在较适宜范围,益生菌对COD无显著影响。5mg·L-1芽孢杆菌和1mg·L-1芽孢杆菌 10mg·L-1光合细菌能显著提高育苗成活率。在实验中,1mg·L-1芽孢杆菌 10mg·L-1光合细菌的综合效果最佳,芽孢杆菌和光合细菌具有协同作用。     

产乳酸芽孢杆菌对对虾养殖水体水质的影响

在温度(28±1)℃、盐度28下,将产乳酸芽孢杆菌制剂添加到养殖10尾凡纳滨对虾、容水200L的0.3m~3室内玻璃钢桶中,使芽孢杆菌终密度为10~4、10~5、10~6 cfu/mL,以无益生菌添加组为对照组。定期测定养殖水体中的氨氮、亚硝酸盐氮和硝酸盐氮的含量,以及总异养菌、弧菌和芽孢杆菌的数量。试验结果显示,试验结束时,添加芽孢杆菌各试验组养殖水体中氨氮、亚硝酸盐氮和弧菌的含量显著低于对照组(P0.05);添加高密度芽孢杆菌试验组(10~6 cfu/mL)养殖水体中总异养菌的数量显著高于对照组(P0.05);添加芽孢杆菌对养殖水体中的硝酸盐氮含量未产生显著的影响(P0.05);添加芽孢杆菌后养殖水体中的芽孢杆菌数量在第4d后会出现下降趋势。试验还发现,在养殖后期初次投入芽孢杆菌时会引起养殖水体中氨氮含量的短期升高。试验结果表明,芽孢杆菌能改善对虾养殖水体水质,可作为益生菌用于对虾养殖中。     

哈氏弧菌文蛤分离株WG1702培养条件优化研究

于2007年9月在江苏吕泗文蛤发病高峰期,从发病文蛤体内和养殖水体中分离到1株哈氏弧菌WG1702,培养条件优化试验结果表明,菌株WG1702在32 h时达到生长高峰,最适温度为28℃,最佳初始pH为7,最佳盐度为40,最佳装液量为60 ml/150 ml,促进生长的金属离子是NH4+、Fe2+、Fe3+,最佳C、N源为葡萄糖和蛋白胨;pH、温度、不同菌龄正交试验证明最佳组合为pH 7.11,温度28℃,菌龄24 h,通过方差分析结果可以看出三因素中pH对生长影响显著(P<0.05)。     

碳氮营养和培养条件对芽孢杆菌(Bacillus sp.) A4生长的影响

芽孢杆菌(Bacillus sp.) A4是一株具有溶甲藻能力的菌株，为探究营养条件与培养条件对A4生长的影响，明确在多因素共同作用下菌株的生长特性，先以单因素方法比较不同碳、氮营养因子对其生长的影响，再以Plackett-Burman方法综合比较碳源、氮源、pH、接种菌量、温度、转速、装液量等因子对其生长的协同影响效应。结果显示，A4菌对有机碳源玉米浆和有机氮源大豆蛋白利用效果最好，培养24 h后菌量分别达到3.58×108、3.19×108 CFU/ml。各因子的重要性排序依次为大豆蛋白、温度、玉米浆、转速、接种菌量、pH、装液量，且大豆蛋白和温度对A4菌的生长影响显著(P<0.05)。研究表明，培养条件对菌株生长调控也有重要意义，在评估相关因素对菌株生长或生态功能的影响时，须将营养条件和培养条件协同分析。     

Lactic acid bacteria effective for regulating the growth of contaminant bacteria during the fermentation of <Emphasis Type="Italic">Undaria pinnatifida</Emphasis> (Phaeophyta)

ABSTRACT:   Lactic acid fermentation of seaweed is a recent topic and quite limited information is available on culture conditions. To know the suitable strains for use as a starter culture for seaweed fermentation, 14 lactic acid bacteria (LAB) strains, including 11 species, were tested in culture conditions prepared with or without salt. A commercial product of Undaria pinnatifida powder was used as a substrate for fermentation without sterilizing. Starter-suitability of the LAB strains was assessed from their predominance after culture. Among the tested strains, Lactobacillus brevis, Lactobacillus plantarum, Lactobacillus casei and Lactobacillus rhamnosus showed high (>90%) predominance in their cultures, while control cultures prepared without inoculation of LAB did not show any detectable growth of acid producing bacteria and spoiled. A total of 102 strains not showing acid producing activity were isolated and characterized from spoiled cultures, and all the isolates were observed as Bacillus strains, including 64 strains (62.7%) and 16 strains (15.7%) of a Bacillus cereu s-related and B. fusiformis -related species, respectively. The Undaria substrate before fermentation contained culturable microorganisms at 1.4–3.1 × 10² CFU/g, but the Bacillus cereu s-related strain was not a major composition, suggesting a concern of selective growth of the Bacillus cereu s-related strain during the spoiled fermentation.     

唾液乳杆菌NF19的液体发酵条件研究

从舟山海域海水中筛选到一株海洋细菌NF19,经16S rDNA分子鉴定和API鉴定结果显示菌株NF19为唾液乳杆菌属(L.salivarius)的一个种,将序列上传至NCBI获得GenBank登录号为FJ405036。通过对菌株NF19进行发酵条件优化及正交试验,得到菌株NF19的最佳发酵条件为:初始pH7.2,接种量为8%,在120 r/min下20℃培养48 h。此条件下发酵液有最佳抑菌效果。     

光合细菌FP04的筛选及对中华鳖养殖水质和生产效益的影响

从入海排污口污泥中筛选到一株有较强净化养殖水质能力的光合细菌,初步鉴定其为红螺菌,命名为FP04。实验室试验表明,FP04能有效降低养殖污水的CODcr含量。在中华鳖的养殖水体中加入不同浓度的FP04,结果与对照组比较,FP04可显著降低养殖水体中的氨氮和化学需氧量,并可减少甲鱼病害发生,提高甲鱼养殖成活率和增重率,其中以养殖水体中FP04浓度为9×104CFU/mL的效果最好,成活率和增重率分别提高14%和31%。     

Infectious pancreatic necrosis virus of salmonids: biological and antigenic features of a pathogenic strain and of a non-pathogenic variant selected in RTG-2 cells

Abstract. A non-pathogenic cell culture adapted variant was obtained from a normally pathogenic strain of IPN virus (Sp type) after several passages in RTG-2 cells. This cell culture modified (CCA) strain was compared with the original wild (W) strain in various tests. Cross neutralization tests showed no obvious antigenic difference between the two. However the CCA strain was neutralized by a 1:5000 dilution of normal trout serum whereas the W strain was not. In RTG-2 cells, CCA strain gave both large and small plaques, whereas the W strain gave only small ones. Both virus strains were heat sensitive and labile to cyclic freezing and thawing. The CCA virus was more stable in storage at 4°C under different pH conditions and its growth in RTG-2 cells was more rapid. The rt (supraoptimal temperature at which viral yield is depressed by 90%) appeared to be 19-20°C for the CCA strain and 18-19°C for the W strain. Pre-treatment of RTG-2 cells with ultraviolet inactivated CCA virus could inhibit growth of W virus, but had no effect on replication of homologous virus.     

Dietary yeast culture supplementation improves initial rearing of McConaughy strain rainbow trout

The addition of a yeast culture food supplement (DVAqua from Diamond V. Mills, Cedar Rapids, IA, USA) was evaluated during feeding of McConaughy strain rainbow trout Oncorhynchus mykiss. Two trials were run consecutively, starting with initial feeding, and were replicated over 2 years. The yeast culture was added to commercial trout starter at levels of 0.125 and 0.25 g kg^?1. The number of rainbow trout that died during the first 4 weeks of rearing was significantly reduced in the tanks receiving either of the yeast culture‐supplemented diets compared with the tanks receiving a control starter diet. Ending tank weights and gain were significantly and consistently greater in the tanks fed the 0.25 g kg^?1 yeast culture diet. Individual fish weights were also significantly heavier in the fish‐fed yeast culture containing diets in each year of the study at the end of 27 days. In the trials from 4 to 8 weeks postinitial feeding, mortality rates were significantly less only in the tanks receiving 0.25 g kg^?1 yeast culture supplementation. Individual fish lengths and weights at the end of the second trial were significantly greater in the tanks receiving either level of yeast culture compared with the control diet only in 1 year. Total tank weights and feed conversion were not significantly different between any of the treatments. The inclusion of DVAqua yeast culture provided significant increases in trout survival and growth through the first 4 weeks of feeding, with continued benefits for the next 4 weeks.     

Relationships between intracellular bacteria and the bivalve killer dinoflagellate Heterocapsa circularisquama (Dinophyceae)

ABSTRACT: Relationships between the growths of five different clonal strains of Heterocapsa circularisquama and intracellular bacteria were investigated using culture experiments. Although each H. circularisquama strain culture was established from one cell by repeated and careful washings with micropipettes, intracellular and extracellular bacteria in each strain culture were still observed under a epifluorescence microscope using the DAPI (4',6-diamidino-2-phenylindole) staining technique. The extracellular bacteria are derived, presumably, from the inside of algal cells after the death and collapse of algal cells in culture. Using an electron microscope, bacteria were constantly observed in the cytoplasm and food vacuoles of H. circularisquama cells. The growth of five algal strains containing bacteria was compared with that of a bacteria-free strain using culture experiments under combined conditions of five different light intensities and five different strengths of culture medium. Bacteria showed no significant effect on the growth or survival of the algal cells. During the algal exponential growth phase, the intracellular bacterial cell numbers per algal cell decreased, whereas the total bacterial cell density in each algal culture increased. Final cell yield (total number) of the intracellular and extracellular bacteria varied considerably according to the algal strains. These results suggest that the intracellular bacteria of H. circularisquama grow or survive depending on the host alga, and that the alga can grow independently.     

An evaluation of DVAqua®, a fully‐fermented yeast culture,during long‐term hatchery rearing of McConaughy strain rainbow trout

The addition of a proprietary, fully‐fermented yeast Saccahromyces cerevisiae culture supplement (DVAqua^®, Diamond V Mills, Cedar Rapids, IA, USA) was evaluated during long‐term feeding of McConaughy strain rainbow trout Oncorhynchus mykiss. Beginning at initial feeding and continuing for 408 days of hatchery rearing, the trout received either a commercially‐manufactured feed or the same feed containing 0.125 g kg^?1 DVAqua. This study was conducted at a production level as part of normal (real‐world) hatchery operations, with the fish periodically inventoried and moved into different rearing units. Although no rearing‐tank replication occurred during the first 54 days of feeding, multiple tanks and raceways were used thereafter. Fish in rearing units receiving DVAqua supplementation exhibited less mortality, particularly during the earlier rearing stages. During the final 177 days of rearing in six raceways, DVAqua‐fed McConaughy strain trout were significantly larger and had a significantly improved feed conversion ratio. The overall feed conversion ratio for the entire duration of the study was 1.17 in the fish receiving DVAqua supplementation compared to 1.21 in the control group. Despite the limitations of this study, the use of DVAqua is recommended for McConaughy strain rainbow trout and other less‐domesticated, more difficult‐to‐rear salmonids.     

熊本牡蛎无嵴和多嵴品系生产性状比较

为了评估熊本牡蛎左壳放射嵴有无对其生产性能的影响,于2013年5月,以熊本牡蛎湛江群体子一代作为基础群体,筛选出无嵴及多嵴品系,采用繁殖生物学方法,比较了两个品系的表型性状差异。结果显示,野生群体中,多嵴亲本规格略大于无嵴亲本,鲜重、壳重及怀卵量均显著大于无嵴亲本;两个品系的受精孵化参数、D型幼虫大小及变态规格彼此间无显著差异;多嵴品系存活力及生长潜力在幼虫及养成期间显著高于无嵴品系。中间育成期间,湛江及大风江牡蛎养殖区环境对两品系生长性状造成了一定程度影响,表现出明显的环境效应,但是对存活力尚未造成影响。熊本牡蛎左壳放射嵴无相对于有是显性性状,主要受到一对显性基因控制,而多嵴性状是隐性性状,可以稳定遗传。研究为熊本牡蛎多嵴品系培育及其左壳放射嵴遗传机制奠定了坚实的理论基础与实践经验。