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CN103667109B - A kind of Rhodobacter capsulatus and application thereof

Publication number

CN103667109B

CN103667109B CN201310501364.3A CN201310501364A CN103667109B CN 103667109 B CN103667109 B CN 103667109B CN 201310501364 A CN201310501364 A CN 201310501364A CN 103667109 B CN103667109 B CN 103667109B

Authority

CN

China

Prior art keywords

rhodobacter

fermented liquid

macroalgae

fermenter

pond

Prior art date

2013-10-23

Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)

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• Micro-Organisms Or Cultivation Processes Thereof (AREA)

本发明公开了一株荚膜红细菌以及应用，本发明提供了荚膜红细菌BN-W-6，它的保藏编号为CGMCC？No.8311，属于光合细菌，经过扩大培养后加入能量源，使其发酵获得发酵产物，发酵产物中包含叶绿素a、类胡萝卜素、酮式-类胡萝卜素、氢化酶和接触酶等。该菌的发酵产物在池塘中，产生的生物酶类，可以使大型藻类逐渐分解，叶绿素a和类胡萝卜素等产物能够吸收大型藻类生长所必须的光照波段使大型藻类萎缩、聚团、直至死亡，并且大型藻类死亡后不影响底质，从而达到控制藻类泛滥生长且不影响海参正常生长的目的。本发明提供的荚膜红细菌发酵液可以抑制和去除大型藻类，从而提高养殖密度，改善了养殖环境，达到提高水生生物池塘养殖产量的目的。The invention discloses a strain of capsulated rhodobacter and its application. The invention provides capsulated rhodobacter BN-W-6, whose preservation number is CGMCC? No.8311, belongs to photosynthetic bacteria. After expanded culture, energy source is added to make it ferment to obtain fermentation products. The fermentation products include chlorophyll a, carotenoids, keto-carotenoids, hydrogenase and catalase, etc. The fermentation product of the bacteria is in the pond, and the biological enzymes produced can gradually decompose the macroalgae, and the products such as chlorophyll a and carotenoids can absorb the light band necessary for the growth of the macroalgae, causing the macroalgae to shrink, agglomerate, and even die , and the death of large algae will not affect the substrate, so as to achieve the purpose of controlling the growth of algae flooding without affecting the normal growth of sea cucumbers. The capsulated rhodobacter fermented liquid provided by the invention can inhibit and remove large algae, thereby increasing the breeding density, improving the breeding environment, and achieving the purpose of increasing the production of aquatic organisms in ponds.

技术领域technical field

本发明涉及一株荚膜红细菌及其应用。The invention relates to a strain of capsulated rhodobacter and its application.

背景技术Background technique

刺参养殖作为黄渤海海域经济的支柱产业，在当地经济发展、促进渔民就业和增加渔民收入等方面有着举足轻重的意义。池塘中大型藻类的泛滥一直是刺参养殖户的心病，刺参是黄渤海海域主要养殖的水产品之一，大型藻类的疯狂生长极大地阻碍了该养殖业的高效发展，甚至威胁到现有产业的生存。大型藻类的繁殖对刺参有极大的影响，大型藻类占据了刺参的生存空间，刺参活动空间减少，还有很多被大型藻类缠绕其中，有的因为不能活动而死亡，有的在人工捞草的过程中被扔掉，投苗量减少，对产量是一个极大的冲击。再则大型藻类在池底的死亡，将释放出大量的氨氮和亚硝酸盐，使刺参的养殖环境迅速恶化，可能会引起大批量的死亡。As a pillar industry in the economy of the Yellow Sea and the Bohai Sea, sea cucumber farming plays a pivotal role in local economic development, promotion of fishermen's employment, and increase of fishermen's income. The flooding of macroalgae in ponds has always been the heart disease of sea cucumber farmers. Sea cucumber is one of the main aquatic products cultured in the Yellow Sea and Bohai Sea. The crazy growth of macroalgae has greatly hindered the efficient development of the aquaculture industry, and even threatened the existing industry survival. The reproduction of large algae has a great impact on the sea cucumber. The large algae occupy the living space of the sea cucumber, the activity space of the sea cucumber is reduced, and many of them are entangled by the large algae. The weeds are thrown away in the process of fishing, and the amount of seedlings thrown is reduced, which has a great impact on the yield. Furthermore, the death of large algae at the bottom of the pool will release a large amount of ammonia nitrogen and nitrite, which will rapidly deteriorate the breeding environment of sea cucumbers and may cause a large number of deaths.

水产领域中的对于预防(抑制)大型藻类研究相对薄弱。这主要是因为大型藻类的类型和生长与普通的养殖水产品存在一定不同，不可控因素较多，很难准确进行除去。现在生产中较为常见的大型藻类就有6，7种之多，每种的生长时间、繁殖方式、适宜条件都不尽相同，有很多藻类在一天时间就长满整个池塘，对刺参的生长极其不利。在人工除草的过程中，还会有一部分的刺参卷在藻类中一并扔掉。有的养殖户在无奈的情况下使用了化学药物，虽可以暂时的除去藻类，但是这样不仅影响了池塘中刺参饵料的生长，对刺参也有较高毒性，同时富集的化学药物也会影响最终的消费者。Research on the prevention (inhibition) of macroalgae in the field of aquatic products is relatively weak. This is mainly because the type and growth of macroalgae are somewhat different from those of ordinary farmed aquatic products, and there are many uncontrollable factors, which make it difficult to remove them accurately. There are as many as 6 or 7 types of large algae that are relatively common in production now, each with different growth time, reproduction mode, and suitable conditions. Many algae can fill the entire pond in one day, which is harmful to the growth of sea cucumbers. Extremely unfavorable. In the process of artificial weeding, some sea cucumber rolls will be thrown away in the algae. Some farmers use chemical drugs in desperation. Although algae can be temporarily removed, this not only affects the growth of sea cucumber bait in the pond, but also has high toxicity to sea cucumbers. At the same time, the enriched chemical drugs will also affect the final consumer.

大型藻类的重复打捞是养殖户在刺参养殖中的一笔不小的开支。捞草工人每人每天接近150元，100亩的刺参养殖池塘，10个工人需要10天左右才能把池塘的大型藻类全部捞净，而在一个月之后，大型藻类又开始影响刺参生存环境。这样算下来，每年的人工除草费用就接近6万。Repeated salvage of macroalgae is a considerable expense for farmers in sea cucumber cultivation. Weeding workers cost close to 150 yuan per person per day. In a 100-acre japonicus breeding pond, 10 workers need about 10 days to remove all the large algae in the pond. After one month, the large algae began to affect the living environment of japonicus. . Calculated in this way, the annual cost of manual weeding is close to 60,000.

光合细菌(PhotosyntheticBacteria，缩写PSB)是水域微生物中的一种，是地球上最早出现的具有原始光能合成系统的原核生物，它广泛分布在各类水体的厌氧层中。现已知光合细菌存在着色杆菌科、外硫红螺菌科、紫色非硫细菌、绿色硫细菌、多细胞丝状绿细菌、螺旋杆菌科、含细菌叶绿素的专性好氧菌等7大群公约50个属。光合细菌作为自然界中重要的微生物类群，广泛存在于自然界的水田、湖泊、江河、海洋、活性污泥及土壤中，生命力极强、营养要求低、生长繁殖快、无毒害性等特点。Photosynthetic bacteria (Photosynthetic Bacteria, abbreviated as PSB) is a kind of microorganism in water area, and it is the earliest prokaryote with primitive photosynthesis system on the earth, which is widely distributed in the anaerobic layer of various water bodies. It is known that there are seven major groups of photosynthetic bacteria including Chromobacteriaceae, Exothorhodospiraceae, purple non-sulfur bacteria, green sulfur bacteria, multicellular filamentous green bacteria, Helicobacteriaceae, and obligate aerobic bacteria containing bacteriochlorophyll. 50 genera. As an important microbial group in nature, photosynthetic bacteria widely exist in paddy fields, lakes, rivers, oceans, activated sludge and soils in nature. They have the characteristics of strong vitality, low nutritional requirements, fast growth and reproduction, and non-toxicity.

发明内容Contents of the invention

本发明的目的是提供一株荚膜红细菌及其应用。具体说，用于水生生物养殖池塘中去除大型藻类。采用本发明的方法养殖池塘水生生物，可使水生生物产量显著增加，其中尤为适宜的水生生物为刺参。The purpose of the present invention is to provide a strain of capsulated rhodobacter and its application. Specifically, it is used to remove macroalgae in aquatic organism breeding ponds. Adopting the method of the invention to cultivate pond aquatic organisms can significantly increase the yield of aquatic organisms, wherein the particularly suitable aquatic organisms are sea cucumbers.

为了达到上述目的，本发明提供一株荚膜红细菌(Rhodobactercapsulatus)BN-W-6，保藏编号为CGMCCNo.8311。In order to achieve the above purpose, the present invention provides a strain of rhodobacter capsulatus (Rhodobacter capsulatus) BN-W-6, the preservation number of which is CGMCC No.8311.

本发明还保护了所述荚膜红细菌BN-W-6在生产抑制和去除藻类菌剂中的应用，用于大型藻类。其中，所述藻类为浒苔、刚毛藻、硬毛藻或者石莼等。The present invention also protects the application of the capsulated rhodobacter BN-W-6 in the production of an agent for inhibiting and removing algae, which is used for large algae. Wherein, the algae is Enteromorpha, Cladophora, Claridioides or Ulva and the like.

本发明荚膜红细菌(Rhodobactorcapsulatus)BN-W-6，可用于制备得到的发酵液。用于抑制和去除藻类的菌剂。The rhodobactor capsulatus BN-W-6 of the present invention can be used to prepare the obtained fermentation liquid. Inoculant for inhibiting and removing algae.

利用所述荚膜红细菌BN-W-6的发酵液的方法为，发酵所述荚膜红细菌BN-W-6，得到发酵液。The method of using the fermented liquid of Rhodobacter capsula BN-W-6 is to ferment the Rhodobacter capsulated BN-W-6 to obtain fermented liquid.

优选方式下，首先将菌浓度扩繁达到10⁹-10¹¹cfu/ml的荚膜红细菌BN-W-6菌液，加入营养物后进行发酵。发酵条件优选为：温度控制在25-30℃，通气流速为2-4L/min，光照强度为1000-3000Lux，发酵24-36h。其中，所述营养物是葡萄糖、蔗糖、红糖或混合构成的营养物；按照1000ml所述菌液加入0.03g营养物的比例加入。In a preferred manner, firstly, the rhodobacter capsular BN-W-6 bacterial liquid with a bacterial concentration of 10 ⁹ -10 ¹¹ cfu/ml is multiplied, and then fermented after adding nutrients. The fermentation conditions are preferably as follows: the temperature is controlled at 25-30° C., the ventilation flow rate is 2-4 L/min, the light intensity is 1000-3000 Lux, and the fermentation is 24-36 hours. Wherein, the nutrient is glucose, sucrose, brown sugar or a mixed nutrient; it is added according to the ratio of 0.03g nutrient to 1000ml of the bacterial solution.

所述发酵液的使用方法为，将所述发酵液按照1.5-3L/亩·米，投放入水生生物养殖池塘。其中，所述发酵液利用沉降剂均匀投放到水生生物养殖池塘中。The method of using the fermented liquid is as follows: put the fermented liquid into aquatic organism breeding ponds at the rate of 1.5-3 L/mu·m. Wherein, the fermented liquid is evenly put into the aquatic organism breeding pond by using a sedimentation agent.

本发明还保护了上述发酵液在水生生物养殖池塘抑制和去除大型藻类中的应用。应用方法包括：将所述发酵液按照1.5-3L/亩·米，投放入水生生物养殖池塘的步骤。The present invention also protects the application of the above-mentioned fermented liquid in inhibiting and removing large algae in aquatic organism culture ponds. The application method includes: putting the fermented liquid into aquatic organism breeding ponds at a rate of 1.5-3 L/mu·m.

所述发酵液尤其适用于水生生物养殖池塘中应用，应用方式，将所述发酵液按照1.5-3L/亩·米，投放入水生生物养殖池塘的步骤。The fermented liquid is especially suitable for application in aquatic organism breeding ponds. The application method is the step of putting the fermented liquid into the aquatic organisms cultivating pond at a rate of 1.5-3 L/mu·m.

本发明提供的荚膜红细菌BN-W-6，属于光合细菌，该细菌经过扩大培养后加入能量源(如葡萄糖、蔗糖或红糖等)，使其发酵获得发酵产物，发酵产物中包含叶绿素a、类胡萝卜素、酮式-类胡萝卜素、氢化酶和接触酶等。该细菌的发酵产物在池塘中，产生的生物酶类，可以使大型藻类逐渐分解，叶绿素a和类胡萝卜素等产物能够吸收大型藻类生长所必须的光照波段使大型藻类萎缩、聚团、直至死亡，并且大型藻类死亡后不影响底质。The capsulated Rhodobacter BN-W-6 provided by the present invention belongs to photosynthetic bacteria. After the bacteria are expanded and cultivated, an energy source (such as glucose, sucrose or brown sugar, etc.) is added to ferment it to obtain a fermentation product, which contains chlorophyll a , carotenoids, keto-carotenoids, hydrogenases and catalases, etc. The fermentation product of the bacteria in the pond produces biological enzymes that can gradually decompose the macroalgae, and products such as chlorophyll a and carotenoids can absorb the light band necessary for the growth of the macroalgae, causing the macroalgae to shrink, agglomerate, and even die , and the death of macroalgae does not affect the substrate.

本发明提供的抑制和去除养殖池塘大型藻类的菌剂可以改善养殖池塘底质微生态环境，降低底质水层氨氮和亚硝酸盐，大型藻类生长所需要的固氮和固碳作用降低，达到抑制大型藻类生长的要求。The bacterial agent for inhibiting and removing macroalgae in aquaculture ponds provided by the invention can improve the bottom micro-ecological environment of aquaculture ponds, reduce ammonia nitrogen and nitrite in the bottom water layer, reduce the nitrogen fixation and carbon fixation required for the growth of macroalgae, and achieve inhibition Requirements for the growth of macroalgae.

本发明提供的抑制和去除养殖池塘大型藻类的菌剂可以进一步提高池塘养殖水生生物的产量，提高养殖生物的成活率，大规模生产抑制和去除养殖池塘大型藻类的菌剂，可以挽救养殖户损失，增加养殖户收入等方面有着举足轻重的意义。本发明尤其适用于黄渤海海域。The bacterial agent for inhibiting and removing macroalgae in cultured ponds provided by the present invention can further increase the output of aquatic organisms cultured in ponds, improve the survival rate of cultured organisms, produce the microbial agent for inhibiting and removing macroalgae in cultured ponds on a large scale, and save the loss of farmers , It is of great significance to increase the income of farmers. The present invention is especially applicable to the sea areas of the Yellow Sea and the Bohai Sea.

具体实施方式detailed description

以下实施例便于更好的理解本发明，但并不限定本发明，下述实施例中的实验方法，如无特殊说明，均为常规方法。下述实施例中所用的试验材料，如无特殊说明，均为自常规生化试剂商店购买得到的。The following examples facilitate a better understanding of the present invention, but do not limit the present invention. The experimental methods in the following examples, unless otherwise specified, are conventional methods. The test materials used in the following examples, unless otherwise specified, were purchased from conventional biochemical reagent stores.

一、菌株BN-W-6的获得1. Obtaining of strain BN-W-6

利用富集和选择鉴别培养技术，从海参肠道分离耐受高浓度氨氮、亚硝酸盐、并能抑制大型藻类生长的光合细菌。得到一株菌株，命名为BN-W-6。Photosynthetic bacteria that tolerate high concentrations of ammonia nitrogen and nitrite and can inhibit the growth of macroalgae were isolated from the intestinal tract of sea cucumbers using enrichment and selective differential culture techniques. A strain was obtained and named as BN-W-6.

二、菌株BN-W-6的鉴定2. Identification of strain BN-W-6

1、形态特征1. Morphological characteristics

革兰氏阴性菌；Gram-negative bacteria;

菌体呈短杆或卵形，大小约为(0.5-1.2)um×(1.8-2.3)um；The bacteria are short rod or oval, with a size of about (0.5-1.2)um×(1.8-2.3)um;

菌落在光照条件下呈光滑、圆润、突起；The colonies are smooth, round and protruding under light conditions;

菌落在光照条件下为红色，在黑暗条件下为无色。Colonies are red under light conditions and colorless under dark conditions.

2、分子特征2. Molecular characteristics

16SrDNA的测序结果如序列表的序列1所示，与GENBANKACCESSIONNO.FJ866782.1的荚膜红细菌的序列相似性为99％。The sequencing result of 16SrDNA is shown as sequence 1 in the sequence table, and the sequence similarity with GENBANKACCESSIONNO.FJ866782.1 of Rhodobacter capsulata is 99%.

综合形态特征和分子特征，参照《伯杰氏系统细菌学手册》(第9版)和东秀珠等编著的《常见细菌系统鉴定手册》(第1版)，菌株BN-W-6属于荚膜红细菌属(Rhodobactercapsulatus)。Comprehensive morphological and molecular characteristics, referring to "Bergey's Handbook of Systematic Bacteriology" (9th edition) and "Handbook of Common Bacterial System Identification" (1st edition) edited by Dong Xiuzhu et al., strain BN-W-6 belongs to capsular Rhodobacter capsulatus.

三、菌株BN-W-6的鉴定保藏3. Identification and preservation of strain BN-W-6

荚膜红细菌(Rhodobactercapsulatus)BN-W-6，简称荚膜红细菌BN-W-6，已于2013年10月9日于中国微生物菌种保藏管理委员会普通微生物中心保藏(简称CGMCC；地址：中国科学院微生物研究所；邮编100101)，保藏编号为CGMCCNo.8311。Rhodobacter capsulatus BN-W-6, referred to as Rhodobacter capsulatus BN-W-6, has been preserved in the General Microbiology Center of China Committee for Microbial Culture Collection (CGMCC for short; address: Institute of Microbiology, Chinese Academy of Sciences; Zip code 100101), the deposit number is CGMCCNo.8311.

四、荚膜红细菌BN-W-6的发酵成份及工艺条件的优化4. Optimization of fermentation components and process conditions of Rhodobacter capsularis BN-W-6

通过单因素及正交试验，确定如下培养基组成参数：Through single factor and orthogonal experiments, the following medium composition parameters were determined:

葡萄糖为培养基中的最佳碳源，添加0.02-0.05％时可得到最高生物量和发酵产物。Glucose is the best carbon source in the medium, and the highest biomass and fermentation products can be obtained when adding 0.02-0.05%.

蛋白胨为培养基中的氮源，添加量为0.5％；Peptone is the nitrogen source in the medium, and the addition amount is 0.5%;

酵母浸粉为培养基中的辅助氮源，添加量为0.1％Yeast extract powder is an auxiliary nitrogen source in the medium, and the addition amount is 0.1%

磷酸高铁作用可以提高菌液的浓度。The action of ferric phosphate can increase the concentration of the bacterial solution.

采用250ml摇瓶，装液量80ml，培养温度25-30℃，Ph6-8，培养时间24-36h。A 250ml shaker flask is used, the liquid volume is 80ml, the culture temperature is 25-30°C, Ph6-8, and the culture time is 24-36h.

采用上述优化参数，荚膜红细菌BN-W-6的生物量达到8x10¹¹ä¸ª/ml，菌体产酶量、产类胡萝卜素量达到最高值。Using the above optimized parameters, the biomass of Rhodobacter capsularis BN-W-6 reached 8x10 ¹¹ cells/ml, and the amount of enzymes and carotenoids produced by the bacteria reached the highest values.

五、抑制和去除养殖池塘大型藻类菌剂的制备5. Preparation of bacterial agents for inhibiting and removing macroalgae in culture ponds

将菌浓度扩繁达到10⁹-10¹¹cfu/ml的荚膜红细菌BN-W-6菌液，加入营养物进行发酵。发酵产物即为抑制和去除养殖池塘大型藻类菌剂。The capsulated rhodobacter BN-W-6 bacterial liquid whose bacterial concentration reaches 10 ⁹ -10 ¹¹ cfu/ml is multiplied, and nutrient is added for fermentation. The fermentation product is the agent for inhibiting and removing macroalgae in the culture pond.

发酵可以选用适宜方式，优选方式下，发酵条件为：温度控制在25-30℃，空气通气流速为2-4L/min，光照强度为1000-3000Lux，发酵24-36h。Fermentation can be done in a suitable way. In the preferred way, the fermentation conditions are as follows: the temperature is controlled at 25-30°C, the air flow rate is 2-4L/min, the light intensity is 1000-3000Lux, and the fermentation is 24-36h.

此外，营养物可以是葡萄糖、蔗糖、红糖或其他类营养物。优选方式下，按照1000ml菌液加入0.03g葡萄糖的比例加入。In addition, the nutrient may be glucose, sucrose, brown sugar or other similar nutrient. In a preferred manner, add 0.03 g of glucose to 1000 ml of bacterial solution.

具体实施方式如下：The specific implementation is as follows:

实施例1、抑制和去除养殖池塘大型藻类菌剂的制备Embodiment 1, inhibit and remove the preparation of culture pond macroalgae bacterial agent

步骤1：菌种液体培养基(pH7.0)的制备方法：5g蛋白胨、1g酵母浸粉、0.01g磷酸高铁、0.04gNaHCO₃、0.02gMgSO₄·7H₂O、0.02gK₂HPO₄、0.1gNH₄Cl，用陈海水定容至1000ml；实际应用中，发酵培养基采用6.5-7.5的pH均可。Step 1: Preparation method of bacterial culture medium (pH7.0): 5g peptone, 1g yeast extract powder, 0.01g ferric phosphate, 0.04gNaHCO ₃ , 0.02gMgSO ₄ 7H ₂ O, 0.02gK ₂ HPO ₄ , 0.1gNH ₄ Cl, dilute to 1000ml with old sea water; in practical application, the pH of the fermentation medium can be 6.5-7.5.

步骤2：将菌种液体培养基在120℃下灭菌15～20min。Step 2: Sterilize the strain liquid culture medium at 120° C. for 15-20 minutes.

步骤3：采用200L发酵罐，并对发酵罐进行如下改造：(1)在发酵罐底部安装发红色的水下灯(灯光主要提供荚膜红细菌的生长必须的光源作为能源)；(2)在发酵罐下部安装电热恒温控制棒；(3)在发酵罐底部设置两个外接开口，一个作为供给发酵罐的通气口，另一个作为加入培养基、营养物和菌种的开口。Step 3: adopt 200L fermenter, and carry out following transformation to fermenter: (1) install red underwater lamp at the bottom of fermenter (the light mainly provides the necessary light source for the growth of capsular rhodobacter as energy); (2) An electric heating constant temperature control rod is installed at the bottom of the fermenter; (3) Two external openings are arranged at the bottom of the fermenter, one is used as a ventilation port for supplying the fermenter, and the other is used as an opening for adding culture medium, nutrients and strains.

在发酵罐中加入160L液体培养基，然后接种荚膜红细菌BN-W-6，菌种扩增条件：温度控制在28℃，空气通气流速为2L/min，光照强度为1500Lux(1000-3000Lux均可)；Add 160L liquid culture medium in the fermenter, then inoculate Rhodobacter capsularis BN-W-6, bacterial strain expansion condition: temperature is controlled at 28 ℃, air ventilation flow rate is 2L/min, light intensity is 1500Lux (1000-3000Lux both);

步骤4：当发酵罐中的菌浓度达到10⁹-10¹¹cfu/ml时，加入营养物进行发酵24-36h(营养物按照1000ml菌液加入0.03g葡萄糖的比例加入)，温度控制在30℃，空气通气流速为2L/min，光照强度为1000-3000Lux，此次选用1500Lux。Step 4: When the bacterial concentration in the fermenter reaches 10 ⁹ -10 ¹¹ cfu/ml, add nutrients to ferment for 24-36 hours (nutrients are added according to the ratio of adding 0.03g glucose to 1000ml bacterial liquid), and the temperature is controlled at 30°C , the air flow rate is 2L/min, and the light intensity is 1000-3000Lux, 1500Lux is selected this time.

步骤5：收集发酵产物，即为去除养殖池塘大型藻类产物，4℃保藏。Step 5: Collect the fermentation product, which is to remove the macroalgae product in the culture pond, and store it at 4°C.

实施例2、抑制和去除养殖池塘大型藻类菌剂的制备Embodiment 2, inhibit and remove the preparation of culture pond macroalgae bacterial agent

步骤1：菌种液体培养基(pH7.0)的制备方法：5g蛋白胨、1g酵母浸粉、0.01g磷酸高铁、0.04gNaHCO₃、0.02gMgSO₄·7H₂O、0.02gK₂HPO₄、0.1gNH₄Cl，用陈海水定容至1000ml；实际应用中，发酵培养基采用6.5-7.5的pH均可。Step 1: Preparation method of bacterial culture medium (pH7.0): 5g peptone, 1g yeast extract powder, 0.01g ferric phosphate, 0.04gNaHCO ₃ , 0.02gMgSO ₄ 7H ₂ O, 0.02gK ₂ HPO ₄ , 0.1gNH ₄ Cl, dilute to 1000ml with old sea water; in practical application, the pH of the fermentation medium can be 6.5-7.5.

步骤2：将菌种液体培养基在120℃下灭菌15-20min。Step 2: Sterilize the strain liquid culture medium at 120° C. for 15-20 minutes.

步骤3：采用200L发酵罐，并对发酵罐进行如下改造：(1)在发酵罐下部安装电热恒温控制棒；(2)在发酵罐底部设置两个外接开口，一个作为供给发酵罐的通气口，另一个作为加入培养基、营养物和菌种的开口。Step 3: Adopt a 200L fermenter, and carry out the following modification on the fermenter: (1) install an electric heating constant temperature control rod at the lower part of the fermenter; (2) set two external openings at the bottom of the fermenter, one as a vent for supplying the fermenter , and the other serves as an opening for adding culture medium, nutrients and strains.

在发酵罐中加入160L液体培养基，然后接种荚膜红细菌BN-W-6，菌种扩增条件：温度控制在28℃，空气通气流速为2L/min，Add 160L of liquid culture medium to the fermenter, and then inoculate Rhodobacter capsularis BN-W-6, strain amplification conditions: the temperature is controlled at 28°C, and the air flow rate is 2L/min.

步骤4：当发酵罐中的菌浓度达到10⁹-10¹¹cfu/ml时，加入营养物后发酵24-36h(营养物按照1000ml菌液加入0.03g葡萄糖)，温度控制在30℃，空气通气流速为2L/min；Step 4: When the bacterial concentration in the fermenter reaches 10 ⁹ -10 ¹¹ cfu/ml, add nutrients and then ferment for 24-36 hours (add 0.03 g of glucose to 1000 ml of bacterial liquid for nutrients), control the temperature at 30°C, and ventilate with air The flow rate is 2L/min;

步骤5：收集发酵产物，即为去除养殖池塘大型藻类产物，4℃保藏。Step 5: Collect the fermentation product, which is to remove the macroalgae product in the culture pond, and store it at 4°C.

实施例3、抑制和去除养殖池塘大型藻类菌剂的制备Embodiment 3, inhibit and remove the preparation of culture pond macroalgae bacterial agent

步骤1：菌种液体培养基(pH7.0)的制备方法：5g蛋白胨、1g酵母浸粉、0.01g磷酸高铁、0.04gNaHCO₃、0.02gMgSO₄·7H₂O、0.02gK₂HPO₄、0.1gNH₄Cl，用陈海水定容至1000ml；实际应用中，发酵培养基采用6.5-7.5的pH均可。Step 1: Preparation method of bacterial culture medium (pH7.0): 5g peptone, 1g yeast extract powder, 0.01g ferric phosphate, 0.04gNaHCO ₃ , 0.02gMgSO ₄ 7H ₂ O, 0.02gK ₂ HPO ₄ , 0.1gNH ₄ Cl, dilute to 1000ml with old sea water; in practical application, the pH of the fermentation medium can be 6.5-7.5.

步骤2：将菌种液体培养基在120℃下灭菌15-20min。Step 2: Sterilize the strain liquid culture medium at 120° C. for 15-20 minutes.

步骤3：采用200L发酵罐，并对发酵罐进行如下改造：(1)在发酵罐底部安装发红色的水下灯(灯光主要提供荚膜红细菌的生长必须的光源作为能源)；(2)在发酵罐下部安装电热恒温控制棒；(3)在发酵罐底部设置两个外接开口，一个作为供给发酵罐的通气口，另一个作为加入培养基、营养物和菌种的开口。Step 3: adopt 200L fermenter, and carry out following transformation to fermenter: (1) install red underwater lamp at the bottom of fermenter (the light mainly provides the necessary light source for the growth of capsular rhodobacter as energy); (2) An electric heating constant temperature control rod is installed at the bottom of the fermenter; (3) Two external openings are arranged at the bottom of the fermenter, one is used as a ventilation port for supplying the fermenter, and the other is used as an opening for adding culture medium, nutrients and strains.

在发酵罐中加入160L液体培养基，然后接种荚膜红细菌BN-W-6，菌种扩增条件：温度控制在28℃，二氧化碳通气流速为4L/min，光照强度为1500Lux(1000-3000Lux均可)；Add 160L liquid medium to the fermenter, and then inoculate Rhodobacter capsularis BN-W-6. Conditions for strain expansion: the temperature is controlled at 28°C, the ventilation flow rate of carbon dioxide is 4L/min, and the light intensity is 1500Lux (1000-3000Lux both);

步骤4：当发酵罐中的菌浓度达到10⁹-10¹¹cfu/ml时，加入营养物后发酵24-36h(营养物按照1000ml菌液加入0.03g葡萄糖)，温度控制在30℃，二氧化碳通气流速为4L/min，光照强度为1500Lux(1000-3000Lux均可)；Step 4: When the bacterial concentration in the fermenter reaches 10 ⁹ -10 ¹¹ cfu/ml, add nutrients and then ferment for 24-36 hours (add 0.03 g of glucose to 1000 ml of bacterial liquid for nutrients), control the temperature at 30°C, and ventilate with carbon dioxide The flow rate is 4L/min, and the light intensity is 1500Lux (1000-3000Lux is acceptable);

步骤5：收集发酵产物，即为去除养殖池塘大型藻类产物，4℃保藏。Step 5: Collect the fermentation product, which is to remove the macroalgae product in the culture pond, and store it at 4°C.

实施例4、抑制和去除养殖池塘大型藻类菌剂的制备Embodiment 4, inhibit and remove the preparation of culture pond macroalgae bacterial agent

步骤1：菌种液体培养基(pH7.0)的制备方法：5g蛋白胨、1g酵母浸粉、0.01g磷酸高铁、0.04gNaHCO₃、0.02gMgSO₄·7H₂O、0.02gK₂HPO₄、0.1gNH₄Cl，用陈海水定容至1000ml；实际应用中，发酵培养基采用6.5-7.5的pH均可。Step 1: Preparation method of bacterial culture medium (pH7.0): 5g peptone, 1g yeast extract powder, 0.01g ferric phosphate, 0.04gNaHCO ₃ , 0.02gMgSO ₄ 7H ₂ O, 0.02gK ₂ HPO ₄ , 0.1gNH ₄ Cl, dilute to 1000ml with old sea water; in practical application, the pH of the fermentation medium can be 6.5-7.5.

步骤2：将菌种液体培养基在120℃下灭菌15-20min。Step 2: Sterilize the strain liquid culture medium at 120° C. for 15-20 minutes.

步骤3：采用200L发酵罐，并对发酵罐进行如下改造：(1)在发酵罐下部安装电热恒温控制棒；(2)在发酵罐底部设置两个外接开口，一个作为供给发酵罐的通气口，另一个作为加入培养基、营养物和菌种的开口。Step 3: Adopt a 200L fermenter, and carry out the following modification on the fermenter: (1) install an electric heating constant temperature control rod at the lower part of the fermenter; (2) set two external openings at the bottom of the fermenter, one as a vent for supplying the fermenter , and the other serves as an opening for adding culture medium, nutrients and strains.

在发酵罐中加入160L液体培养基，然后接种荚膜红细菌BN-W-6，菌种扩增条件：温度控制在28℃，二氧化碳通气流速为4L/min。Add 160L of liquid culture medium into the fermenter, and then inoculate Rhodobacter capsularis BN-W-6, strain amplification conditions: the temperature is controlled at 28°C, and the flow rate of carbon dioxide ventilation is 4L/min.

步骤4：当发酵罐中的菌浓度达到10⁹-10¹¹cfu/ml时，加入营养物后发酵24-36h(营养物按照1000ml菌液加入0.03g葡萄糖)，温度控制在30℃，二氧化碳通气流速为4L/min。Step 4: When the bacterial concentration in the fermenter reaches 10 ⁹ -10 ¹¹ cfu/ml, add nutrients and then ferment for 24-36 hours (add 0.03 g of glucose to 1000 ml of bacterial liquid for nutrients), control the temperature at 30°C, and ventilate with carbon dioxide The flow rate is 4L/min.

步骤5：收集发酵产物，即为去除养殖池塘大型藻类产物，4℃保藏。Step 5: Collect the fermentation product, which is to remove the macroalgae product in the culture pond, and store it at 4°C.

实施例5、抑制和去除养殖池塘大型藻类菌剂的制备Embodiment 5, inhibit and remove the preparation of culture pond macroalgae bacterial agent

步骤1：菌种液体培养基(pH7.0)的制备方法：10g蛋白胨、2g酵母浸粉、0.1g磷酸高铁、0.04gNaHCO₃、0.02gMgSO₄·7H₂O、0.02gK₂HPO₄、0.1gNH₄Cl，用陈海水定容至1000ml；实际应用中，发酵培养基采用6.5-7.5的pH均可。Step 1: Preparation method of bacterial culture medium (pH7.0): 10g peptone, 2g yeast extract powder, 0.1g ferric phosphate, 0.04gNaHCO ₃ , 0.02gMgSO ₄ 7H ₂ O, 0.02gK ₂ HPO ₄ , 0.1gNH ₄ Cl, dilute to 1000ml with old sea water; in practical application, the pH of the fermentation medium can be 6.5-7.5.

步骤2：将菌种液体培养基在120℃下灭菌15-20min。Step 2: Sterilize the strain liquid culture medium at 120° C. for 15-20 minutes.

步骤3：采用200L发酵罐，并对发酵罐进行如下改造：(1)在发酵罐底部安装发红色的水下灯(灯光主要提供荚膜红细菌的生长必须的光源作为能源)；(2)在发酵罐下部安装电热恒温控制棒；(3)在发酵罐底部设置两个外接开口，一个作为供给发酵罐的通气口，另一个作为加入培养基、营养物和菌种的开口。Step 3: adopt 200L fermenter, and carry out following transformation to fermenter: (1) install red underwater lamp at the bottom of fermenter (the light mainly provides the necessary light source for the growth of capsular rhodobacter as energy); (2) An electric heating constant temperature control rod is installed at the bottom of the fermenter; (3) Two external openings are arranged at the bottom of the fermenter, one is used as a ventilation port for supplying the fermenter, and the other is used as an opening for adding culture medium, nutrients and strains.

在发酵罐中加入160L液体培养基，然后接种荚膜红细菌BN-W-6，菌种扩增条件：温度控制在28℃，空气通气流速为2L/min，光照强度为1500Lux(1000-3000Lux均可)；Add 160L liquid culture medium in the fermenter, then inoculate Rhodobacter capsularis BN-W-6, bacterial strain expansion condition: temperature is controlled at 28 ℃, air ventilation flow rate is 2L/min, light intensity is 1500Lux (1000-3000Lux both);

步骤4：当发酵罐中的菌浓度达到10⁹-10¹¹cfu/ml时，加入营养物后发酵24-36h(按照1000ml菌液加入0.02g蔗糖、0.02gMgSO₄·7H₂O和0.02gK₂HPO₄计算)，温度控制在30℃，空气通气流速为2L/min，光照强度为1500Lux(1000-3000Lux均可)；Step 4: When the bacterial concentration in the fermenter reaches 10 ⁹ -10 ¹¹ cfu/ml, add nutrients and then ferment for 24-36 hours (add 0.02g sucrose, 0.02gMgSO ₄ ·7H ₂ O and 0.02gK ₂ to 1000ml bacterial liquid HPO ₄ calculation), the temperature is controlled at 30°C, the air flow rate is 2L/min, and the light intensity is 1500Lux (1000-3000Lux is acceptable);

步骤5：收集发酵产物，即为去除养殖池塘大型藻类产物，4℃保藏。Step 5: Collect the fermentation product, which is to remove the macroalgae product in the culture pond, and store it at 4°C.

实施例6、抑制和去除养殖池塘大型藻类菌剂的制备Embodiment 6, inhibit and remove the preparation of culture pond macroalgae bacterial agent

步骤1：菌种液体培养基(pH7.0)的制备方法：5g蛋白胨、1g酵母浸粉、0.01g磷酸高铁、用陈海水定容至1000ml；实际应用中，发酵培养基采用6.5-7.5的pH均可。Step 1: The preparation method of the strain liquid medium (pH7.0): 5g peptone, 1g yeast extract powder, 0.01g ferric phosphate, and dilute to 1000ml with old sea water; in practical application, the fermentation medium is 6.5-7.5 pH can be.

步骤2：将菌种液体培养基在120℃下灭菌15-20min。Step 2: Sterilize the strain liquid culture medium at 120° C. for 15-20 minutes.

步骤3：采用200L发酵罐，并对发酵罐进行如下改造：(1)在发酵罐下部安装电热恒温控制棒；(2)在发酵罐底部设置两个外接开口，一个作为供给发酵罐的通气口，另一个作为加入培养基、营养物和菌种的开口。Step 3: Adopt a 200L fermenter, and carry out the following modification on the fermenter: (1) install an electric heating constant temperature control rod at the lower part of the fermenter; (2) set two external openings at the bottom of the fermenter, one as a vent for supplying the fermenter , and the other serves as an opening for adding culture medium, nutrients and strains.

在发酵罐中加入160L液体培养基，然后接种荚膜红细菌BN-W-6，菌种扩增条件：温度控制在28℃，空气通气流速为2L/min，Add 160L of liquid culture medium to the fermenter, and then inoculate Rhodobacter capsularis BN-W-6, strain amplification conditions: the temperature is controlled at 28°C, and the air flow rate is 2L/min.

步骤4：当发酵罐中的菌浓度达到10⁹-10¹¹cfu/ml时，加入营养物后发酵24-36h(按照1000ml菌液加入0.03g蔗糖)，温度控制在30℃，空气通气流速为2L/min；Step 4: When the bacterial concentration in the fermenter reaches 10 ⁹ -10 ¹¹ cfu/ml, add nutrients and then ferment for 24-36 hours (according to 1000ml bacterial liquid, add 0.03g sucrose), the temperature is controlled at 30°C, and the air flow rate is 2L/min;

步骤5：收集发酵产物，即为去除养殖池塘大型藻类产物，4℃保藏。Step 5: Collect the fermentation product, which is to remove the macroalgae product in the culture pond, and store it at 4°C.

实施例7、抑制和去除养殖池塘大型藻类菌剂的制备Embodiment 7, inhibit and remove the preparation of culture pond macroalgae bacterial agent

步骤1：菌种液体培养基(pH7.0)的制备方法：10g蛋白胨、2g酵母浸粉、0.1g磷酸高铁、0.04gNaHCO₃用陈海水定容至1000ml；实际应用中，发酵培养基采用6.5-7.5的pH均可。Step 1: The preparation method of the strain liquid culture medium (pH7.0): 10g peptone, 2g yeast extract powder, 0.1g ferric phosphate, 0.04g NaHCO ₃ Dilute to 1000ml with old sea water; in practical application, the fermentation medium is 6.5 A pH of -7.5 is acceptable.

步骤2：将菌种液体培养基在120℃下灭菌15-20min。Step 2: Sterilize the strain liquid culture medium at 120° C. for 15-20 minutes.

步骤3：采用200L发酵罐，并对发酵罐进行如下改造：(1)在发酵罐底部安装发红色的水下灯(灯光主要提供荚膜红细菌的生长必须的光源作为能源)；(2)在发酵罐下部安装电热恒温控制棒；(3)在发酵罐底部设置两个外接开口，一个作为供给发酵罐的通气口，另一个作为加入培养基、营养物和菌种的开口。Step 3: adopt 200L fermenter, and carry out following transformation to fermenter: (1) install red underwater lamp at the bottom of fermenter (the light mainly provides the necessary light source for the growth of capsular rhodobacter as energy); (2) An electric heating constant temperature control rod is installed at the bottom of the fermenter; (3) Two external openings are arranged at the bottom of the fermenter, one is used as a ventilation port for supplying the fermenter, and the other is used as an opening for adding culture medium, nutrients and strains.

在发酵罐中加入160L液体培养基，然后接种荚膜红细菌BN-W-6，菌种扩增条件：温度控制在28℃，二氧化碳通气流速为4L/min，光照强度为1500Lux(1000-3000Lux均可)；Add 160L liquid medium to the fermenter, and then inoculate Rhodobacter capsularis BN-W-6. Conditions for strain expansion: the temperature is controlled at 28°C, the ventilation flow rate of carbon dioxide is 4L/min, and the light intensity is 1500Lux (1000-3000Lux both);

步骤4：当发酵罐中的菌浓度达到10⁹-10¹¹cfu/ml时，加入营养物后发酵24-36h(按照1000ml菌液加入0.03g蔗糖计算)，温度控制在30℃，二氧化碳通气流速为4L/min，光照强度为1500Lux(1000-3000Lux均可)；Step 4: When the bacterial concentration in the fermenter reaches 10 ⁹ -10 ¹¹ cfu/ml, add nutrients and ferment for 24-36 hours (calculated based on adding 0.03g sucrose to 1000ml bacterial liquid), control the temperature at 30°C, and ventilate the flow rate of carbon dioxide 4L/min, the light intensity is 1500Lux (1000-3000Lux is acceptable);

步骤5：收集发酵产物，即为去除养殖池塘大型藻类产物，4℃保藏。Step 5: Collect the fermentation product, which is to remove the macroalgae product in the culture pond, and store it at 4°C.

实施例8、抑制和去除养殖池塘大型藻类菌剂的制备Embodiment 8, inhibit and remove the preparation of culture pond macroalgae bacterial agent

步骤1：菌种液体培养基(pH7.0)的制备方法：10g蛋白胨、2g酵母浸粉、0.01g磷酸高铁、0.02gK₂HPO₄、0.01gNH₄Cl，用水定容至1000ml；实际应用中，发酵培养基采用6.5-7.5的pH均可。Step 1: Preparation method of strain liquid medium (pH7.0): 10g peptone, 2g yeast extract powder, 0.01g ferric phosphate, 0.02gK ₂ HPO ₄ , 0.01gNH ₄ Cl, dilute to 1000ml with water; , The pH of the fermentation medium can be 6.5-7.5.

步骤2：将菌种液体培养基在120℃下灭菌15-20min。Step 2: Sterilize the strain liquid culture medium at 120° C. for 15-20 minutes.

步骤3：采用200L发酵罐，并对发酵罐进行如下改造：(1)在发酵罐下部安装电热恒温控制棒；(2)在发酵罐底部设置两个外接开口，一个作为供给发酵罐的通气口，另一个作为加入培养基、营养物和菌种的开口。Step 3: Adopt a 200L fermenter, and carry out the following modification on the fermenter: (1) install an electric heating constant temperature control rod at the lower part of the fermenter; (2) set two external openings at the bottom of the fermenter, one as a vent for supplying the fermenter , and the other serves as an opening for adding culture medium, nutrients and strains.

在发酵罐中加入160L液体培养基，然后接种荚膜红细菌BN-W-6，菌种扩增条件：温度控制在28℃，二氧化碳通气流速为4L/min。Add 160L of liquid culture medium into the fermenter, and then inoculate Rhodobacter capsularis BN-W-6, strain amplification conditions: the temperature is controlled at 28°C, and the flow rate of carbon dioxide ventilation is 4L/min.

步骤4：当发酵罐中的菌浓度达到10⁹-10¹¹cfu/ml时，加入营养物后发酵24-36h(按照1000ml菌液加入0.03g葡萄糖和0.03g蔗糖计算)，温度控制在30℃，二氧化碳通气流速为4L/min。Step 4: When the bacterial concentration in the fermenter reaches 10 ⁹ -10 ¹¹ cfu/ml, add nutrients and then ferment for 24-36 hours (calculated based on adding 0.03g glucose and 0.03g sucrose to 1000ml bacterial solution), and control the temperature at 30°C , the carbon dioxide ventilation flow rate is 4L/min.

步骤5：收集发酵产物，即为去除养殖池塘大型藻类产物，4℃保藏。Step 5: Collect the fermentation product, which is to remove the macroalgae product in the culture pond, and store it at 4°C.

实施例9、抑制和去除养殖池塘大型藻类菌剂的制备Embodiment 9, inhibit and remove the preparation of culture pond macroalgae bacterial agent

步骤1：发酵培养基(pH7.0)的制备方法：取0.03g葡萄糖、5g蛋白胨、1g酵母浸粉、0.04gNaHCO₃、0.02gMgSO₄·7H₂O、0.02gK₂HPO₄、0.1gNH₄Cl和10ml微量元素溶液，用陈海水定容至1000ml；所述微量元素溶液的制备方法如下：取0.5gH₃BO₃、400mgMnSO₄、60mgZnSO₄·7H₂O、180mgNaMoO₄·2H₂O和Ca(NO₃)₂·2H₂O，用水溶解并定容至250ml。实际应用中，发酵培养基采用6.5-7.5的pH均可。Step 1: Preparation method of fermentation medium (pH7.0): Take 0.03g glucose, 5g peptone, 1g yeast extract powder, 0.04gNaHCO ₃ , 0.02gMgSO ₄ 7H ₂ O, 0.02gK ₂ HPO ₄ , 0.1gNH ₄ Cl and 10ml trace element solution, dilute to 1000ml with old sea water; the preparation method of the trace element solution is as follows: take 0.5gH ₃ BO ₃ , 400mgMnSO ₄ , 60mgZnSO ₄ ·7H ₂ O, 180mgNaMoO ₄ ·2H ₂ O and Ca( NO ₃ ) ₂ ·2H ₂ O, dissolved in water and adjusted to 250ml. In practical applications, the pH of the fermentation medium may be 6.5-7.5.

步骤2：将发酵培养基在120℃下灭菌15-20min。Step 2: Sterilize the fermentation medium at 120° C. for 15-20 minutes.

步骤3：采用200L发酵罐，并对发酵罐进行如下改造：(1)在发酵罐底部安装发红色的水下灯(灯光主要提供光合细菌的生长必须的光源作为能源)；(2)在发酵罐下部安装电热恒温控制棒；(3)在发酵罐底部设置两个外接开口，一个作为供给发酵罐二氧化碳的通气口，另一个作为加入发酵培养基和菌种的开口。Step 3: Adopt a 200L fermenter, and carry out the following modification to the fermenter: (1) install a red underwater light at the bottom of the fermenter (the light mainly provides the necessary light source for the growth of photosynthetic bacteria as energy); An electric heating constant temperature control rod is installed at the bottom of the tank; (3) Two external openings are arranged at the bottom of the fermenter, one is used as a vent for supplying carbon dioxide to the fermenter, and the other is used as an opening for adding fermentation medium and strains.

在发酵罐中加入160L发酵培养基，然后接种荚膜红细菌BN-W-6，发酵条件：温度控制在30℃，二氧化碳通气流速为4L/min，光照强度为1500Lux(1000-3000Lux均可)；发酵培养24-36h，收集产物，即为去除养殖池塘大型藻类菌剂，4℃保藏。Add 160L of fermentation medium into the fermenter, and then inoculate Rhodobacter capsularis BN-W-6. Fermentation conditions: temperature is controlled at 30°C, carbon dioxide ventilation flow rate is 4L/min, light intensity is 1500Lux (1000-3000Lux is acceptable) ; After 24-36 hours of fermentation and cultivation, the product is collected, which is to remove macroalgae in the culture pond, and stored at 4°C.

六、菌剂的投放，用于抑制和去除养殖池塘大型藻类以及水生生物养殖的应用。6. The application of bacterial agents to inhibit and remove macroalgae in aquaculture ponds and aquatic organisms.

将上述任一实施例发酵的菌剂按照每亩·米投放发酵产物1.5-3L的比例，利用沉降剂均匀投放到水生生物养殖池塘中。优选实施例中水生生物为刺参。The bacteria agent fermented in any of the above-mentioned examples is put into the aquatic organism breeding pond evenly by using the sedimentation agent at a ratio of 1.5-3L of the fermentation product per mu·m. In a preferred embodiment, the aquatic organism is sea cucumber.

注意投放7-12天内不能进行池塘换水，防止发酵产物浓度降低，失去抑制大型藻类效果。Note that the water in the pond cannot be changed within 7-12 days of putting it in, so as to prevent the concentration of fermentation products from decreasing and losing the effect of inhibiting macroalgae.

具体实施例如下：发现大型藻类生长时，首先制备上述菌剂及获取沉降剂(可购买取得)。计算池塘总面积，按照每亩·米投放发酵产物1.5-3L计算。沉降剂可选用海泥或蛎壳粉。Specific examples are as follows: when macroalgae are found to grow, first prepare the above bacterial agent and obtain a sedimentation agent (available for purchase). To calculate the total area of the pond, it is calculated by adding 1.5-3L of fermentation product per mu·meter. The sedimentation agent can be sea mud or oyster shell powder.

计算池塘总面积，按照每亩·米投放发酵产物1.5-3L计算。具体方法如下池塘面积按照50亩，池塘水深1米，发酵产物使用量为2L/亩·米。To calculate the total area of the pond, it is calculated by adding 1.5-3L of fermentation product per mu·meter. The specific method is as follows: the pond area is 50 mu, the pond water depth is 1 meter, and the fermentation product usage is 2L/mu·meter.

依次进行以下操作：Do the following in order:

(1)将发酵产物100L，倒入大水槽中，并添加入200L-400L的海水。(1) Pour 100L of fermentation product into a large water tank, and add 200L-400L of seawater.

(2)将20kg-40kg海泥放入已经装好发酵产物的水槽中(2) Put 20kg-40kg of sea mud into the tank that has been filled with fermentation products

(3)将发酵产物和海泥搅拌均匀，如搅拌时太粘稠，可以适量再加入200L-400L海水。(3) Stir the fermentation product and sea mud evenly. If it is too thick when stirring, you can add 200L-400L seawater in an appropriate amount.

(4)连续搅动10-15min(4) Stir continuously for 10-15min

(5)稀释后均匀泼洒到有大型藻类的所在池塘水面，让其自然沉底。(5) After dilution, evenly sprinkle it on the water surface of the pond where there are large algae, and let it sink to the bottom naturally.

(6)泼洒后7-12天内不能进行池塘换水，防止发酵产物浓度降低，失去抑制大型藻类效果。(6) Pond water cannot be changed within 7-12 days after splashing to prevent the concentration of fermentation products from decreasing and losing the effect of inhibiting macroalgae.

水温大于8℃时使用，风速小于三级时使用，在傍晚16点到19点投放效果最佳。Use it when the water temperature is higher than 8°C, use it when the wind speed is lower than level 3, and put it in between 16:00 and 19:00 in the evening for the best effect.

投放第4天后，能够发现大型藻类有变黄，萎缩的情况发生，藻类可以由原来的10-20cm，萎缩至7-15cm左右，大一些的可以从50-70cm，萎缩到40-50cm。After the 4th day of feeding, it can be found that the large algae have turned yellow and shrunk. The algae can shrink from the original 10-20cm to about 7-15cm, and the larger ones can shrink from 50-70cm to 40-50cm.

投放第7‐10天后，能够发现大型藻类变黄，萎缩的情况加剧，藻类尖端发黑，效果明显，藻类可以由原来的10‐20cm，萎缩至2‐3cm左右，大一些的可以从50‐70cm，萎缩到10‐20cm。藻类可以由上述实施过程尤其适用于黄渤海海域气候条件下的池塘养殖。通过上述过程，实践中，本发明基于生物技术控制海参池塘大型藻类的方法，能够抑制大型藻类在池塘中的繁殖。After the 7-10th day of placement, it can be found that the large algae turn yellow, the atrophy is intensified, and the tip of the algae turns black. The effect is obvious. The algae can shrink from the original 10-20cm to about 2-3cm. 70cm, shrinking to 10‐20cm. The algae can be cultured in ponds under the climatic conditions of the Yellow Sea and the Bohai Sea by the above-mentioned implementation process. Through the above-mentioned process, in practice, the method of the present invention for controlling macroalgae in sea cucumber ponds based on biotechnology can inhibit the reproduction of macroalgae in the pond.

本发明公开了一株荚膜红细菌以及应用，本发明提供了荚膜红细菌BN-W-6，它的保藏编号为CGMCCNo.8311，属于光合细菌，该细菌经过扩大培养后加入能量源(如葡萄糖、蔗糖或红糖等)，使其发酵获得发酵产物，发酵产物中包含叶绿素a、类胡萝卜素、酮式-类胡萝卜素、氢化酶和接触酶等。该细菌的发酵产物在池塘中，产生的生物酶类，可以使大型藻类逐渐分解，叶绿素a和类胡萝卜素等产物能够吸收大型藻类生长所必须的光照波段使大型藻类萎缩、聚团、直至死亡，并且大型藻类死亡后不影响底质，从而达到控制藻类泛滥生长且不影响海参正常生长的目的。本发明提供的荚膜红细菌可以抑制和去除大型藻类，从而提高了养殖密度，改善了养殖环境，达到提高水生生物池塘养殖的产量The invention discloses a strain of capsulated rhodobacter and its application. The invention provides capsulated rhodobacter BN-W-6, whose preservation number is CGMCCNo.8311, which belongs to photosynthetic bacteria. After the bacteria are expanded and cultivated, an energy source ( Such as glucose, sucrose or brown sugar, etc.), make it ferment to obtain fermented product, which contains chlorophyll a, carotenoid, keto-carotenoid, hydrogenase and catalase etc. The fermentation product of the bacteria in the pond produces biological enzymes that can gradually decompose the macroalgae, and products such as chlorophyll a and carotenoids can absorb the light band necessary for the growth of the macroalgae, causing the macroalgae to shrink, agglomerate, and even die , and the death of large algae will not affect the substrate, so as to achieve the purpose of controlling the growth of algae flooding without affecting the normal growth of sea cucumbers. The capsulated rhodobacteria provided by the invention can inhibit and remove macroalgae, thereby increasing the culture density, improving the culture environment, and increasing the yield of aquatic organism pond culture

以上所述，仅为本发明较佳的具体实施方式，但本发明的保护范围并不局限于此，任何熟悉本技术领域的技术人员在本发明披露的技术范围内，根据本发明的技术方案及其发明构思加以等同替换或改变，都应涵盖在本发明的保护范围之内。The above is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Anyone familiar with the technical field within the technical scope disclosed in the present invention, according to the technical solution of the present invention Any equivalent replacement or change of the inventive concepts thereof shall fall within the protection scope of the present invention.

1.荚膜红细菌(Rhodobactercapsulatus)BN-W-6，其特征在于，保藏编号为CGMCCNo.8311。 1. Rhodobacter capsulatus BN-W-6, characterized in that the preservation number is CGMCCNo.8311. 2.权利要求1所述荚膜红细菌BN-W-6在生产抑制和去除藻类菌剂中的应用，其特征在于，用于大型藻类。 2. the application of the capsulated rhodobacter BN-W-6 of claim 1 in the production of inhibition and removal of algae inoculum, characterized in that it is used for macroalgae. 3.权利要求2所述荚膜红细菌BN-W-6在生产抑制和去除藻类菌剂中的应用，其特征在于，所述藻类为浒苔、刚毛藻、硬毛藻或者石莼。 3. The application of the capsulated rhodobacter BN-W-6 of claim 2 in the production of inhibition and removal of algae inoculants, characterized in that, the algae is Enteromorpha, Cladophora, Clarides or Ulva. 4.一种制备荚膜红细菌BN-W-6的发酵液的方法，其特征在于，发酵权利要求1所述荚膜红细菌BN-W-6，得到发酵液。 4. A method for preparing a fermented liquid of Rhodobacter capsularis BN-W-6, characterized in that, fermenting Rhodobacter capsulated BN-W-6 according to claim 1, obtains fermented liquid. 5.根据权利要求4所述制备荚膜红细菌BN-W-6的发酵液的方法，其特征在于，首先将菌浓度扩繁达到10⁹-10¹¹cfu/ml的荚膜红细菌BN-W-6菌液，加入营养物后进行发酵。 5. The method for preparing the fermented liquid of Rhodobacter capsularis BN-W- ⁶ according to claim ⁴ , is characterized in that, at first the capsulated Rhodobacterium BN-W- W-6 bacterial liquid, fermented after adding nutrients. 6.根据权利要求5所述制备荚膜红细菌BN-W-6的发酵液的方法，其特征在于，发酵条件为：温度控制在25-30℃，通气流速为2-4L/min，光照强度为1000-3000Lux，发酵24-36h； 6. The method for preparing the fermented liquid of Rhodobacter capsularis BN-W-6 according to claim 5, is characterized in that, fermentation condition is: temperature is controlled at 25-30 ℃, and ventilation flow rate is 2-4L/min, light The strength is 1000-3000Lux, fermented for 24-36h; 所述营养物是葡萄糖、蔗糖、红糖或混合构成的营养物；按照1000ml所述菌液加入0.03g营养物的比例加入。 The nutrient is glucose, sucrose, brown sugar or a mixed nutrient; it is added according to the ratio of 0.03g nutrient to 1000ml of the bacterial solution. 7.权利要求4-6任一所述方法制备得到的发酵液。 7. the fermented liquid prepared by the arbitrary described method of claim 4-6. 8.权利要求7所述发酵液的使用方法，其特征在于，该方法是将所述发酵液按照1.5-3L/亩·米，投放入水生生物养殖池塘。 8. The method for using the fermented liquid according to claim 7, characterized in that, the method is to put the fermented liquid into the aquatic organism culture pond according to 1.5-3 L/mu·m. 9.根据权利要求8所述发酵液的使用方法，其特征在于，所述发酵液利用沉降剂均匀投放到水生生物养殖池塘中。 9. The using method of the fermented liquid according to claim 8, characterized in that, the fermented liquid is evenly put into the aquatic organism culture pond by using a settling agent. 10.一种权利要求7所述发酵液在水生生物养殖池塘抑制和去除大型藻类中的应用。 10. the application of a kind of fermented liquid described in claim 7 suppresses and removes macroalgae in aquatic organism culture pond. 11.根据权利要求10所述发酵液在水生生物养殖池塘抑制和去除大型藻类中的应用，其特征在于，该方法包括：将权利要求4-6任一所述发酵液按照1.5-3L/亩·米，投放入水生生物养殖池塘的步骤。 11. The application of the fermented liquid according to claim 10 in the inhibition and removal of macroalgae in aquatic organism culture ponds, characterized in that the method comprises: using the fermented liquid according to any one of claims 4-6 at a rate of 1.5-3L/mu Rice, the step of throwing it into the aquatic organism culture pond. 12.一种权利要求7所述发酵液在水生生物养殖池塘中的应用。 12. an application of the fermented liquid described in claim 7 in aquatic organism culture ponds. 13.根据权利要求12所述发酵液在水生生物养殖池塘中的应用，其特征在于，将权利要求4-6任一所述发酵液按照1.5-3L/亩·米，投放入水生生物养殖池塘的步骤。 13. according to the application of the fermented liquid described in claim 12 in the pond of aquatic organism cultivation, it is characterized in that, according to any one of claim 4-6 fermented liquid according to 1.5-3L/mu·meter, put into pond for aquatic organism culture A step of.

• 2013
  • 2013-10-23 CN CN201310501364.3A patent/CN103667109B/en not_active Expired - Fee Related

Granted publication date: 20160203

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